Author: "Guy Vernet" / Topic: medicine - Searchworks@Jio Institute Digital Library Search Results

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1. Surveillance for Antimicrobial Drug Resistance in Under-Resourced Countries

Author: Guy Vernet, Catherine Mary, Dany M. Altmann, Ogobara Doumbo, Susan Morpeth, Zulfiqar A. Bhutta, and Keith P. Klugman
Subjects: antimicrobial resistance, surveillance, under-resourced countries, diagnostics, Medicine, Infectious and parasitic diseases, RC109-216
Abstract: Antimicrobial drug resistance is usually not monitored in under-resourced countries because they lack surveillance networks, laboratory capacity, and appropriate diagnostics. This accelerating problem accounts for substantial number of excess deaths, especially among infants. Infections particularly affected by antimicrobial drug resistance include tuberculosis, malaria, severe acute respiratory infections, and sepsis caused by gram-negative bacteria. Nonetheless, mapping antimicrobial drug resistance is feasible in under-resourced countries, and lessons can be learned from previous successful efforts. Specimen shipping conditions, data standardization, absence of contamination, and adequate diagnostics must be ensured. As a first step toward solving this problem, we propose that a road map be created at the international level to strengthen antimicrobial resistance surveillance in under-resourced countries. This effort should include a research agenda; a map of existing networks and recommendations to unite them; and a communication plan for national, regional, and international organizations and funding agencies. Download MP3 Length: 1:26
Published: 2014
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2. Coxsackievirus A21, Enterovirus 68, and Acute Respiratory Tract Infection, China

Author: Zichun Xiang, Richard Gonzalez, Zhong Wang, Lili Ren, Yan Xiao, Jianguo Li, Yongjun Li, Guy Vernet, Gláucia Paranhos-Baccalà, Qi Jin, and Jianwei Wang
Subjects: Coxsackievirus A21, enterovirus 68, adults, respiratory tract infection, viruses, China, Medicine, Infectious and parasitic diseases, RC109-216
Abstract: During August 2006–April 2010, in Beijing, China, 2 rare human enterovirus serotypes, coxsackievirus A21 and enterovirus 68, were detected most frequently in human enterovirus–positive adults with acute respiratory tract infections. Thus, during some years, these 2 viruses cause a substantial proportion of enterovirus-associated adult acute respiratory tract infections.
Published: 2012
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3. Novel Human Reovirus Isolated from Children with Acute Necrotizing Encephalopathy

Author: Louise A. Ouattara, Francis Barin, Marie Anne Barthez, Bertrand Bonnaud, Philippe Roingeard, Alain Goudeau, Pierre Castelnau, Guy Vernet, Gláucia Paranhos-Baccalà, and Florence Komurian-Pradel
Subjects: encephalitis, mammalian orthoreovirus 2, reassortant viruses, viruses, research, France, Medicine, Infectious and parasitic diseases, RC109-216
Abstract: For many encephalitis cases, the cause remains unidentified. After 2 children (from the same family) received a diagnosis of acute necrotizing encephalopathy at Centre Hospitalier Universitaire (Tours, France), we attempted to identify the etiologic agent. Because clinical samples from the 2 patients were negative for all pathogens tested, urine and throat swab specimens were added to epithelial cells, and virus isolates detected were characterized by molecular analysis and electron microscopy. We identified a novel reovirus strain (serotype 2), MRV2Tou05, which seems to be closely related to porcine and human strains. A specific antibody response directed against this new reovirus strain was observed in convalescent-phase serum specimens from the patients, whereas no response was observed in 38 serum specimens from 38 healthy adults. This novel reovirus is a new etiologic agent of encephalitis.
Published: 2011
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4. Respiratory Infection with Enterovirus Genotype C117, China and Mongolia

Author: Zichun Xiang, Sosorbaramyn Tsatsral, Chunyan Liu, Linlin Li, Lili Ren, Yan Xiao, Zhengde Xie, Hongli Zhou, Guy Vernet, Pagbajabyn Nymadawa, Kunling Shen, and Jianwei Wang
Subjects: EV-C117,picornavirus, enterovirus, viruses, respiratory tract infection, children, pediatric, Medicine, Infectious and parasitic diseases, RC109-216
Published: 2014
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5. Saffold Cardioviruses of 3 Lineages in Children with Respiratory Tract Infections, Beijing, China

Author: Lili Ren, Richard Gonzalez, Zhengde Xie, Yan Xiao, Yongjun Li, Chunyan Liu, Lan Chen, Qingqing Yang, Guy Vernet, Gláucia Paranhos-Baccalà, Qi Jin, Kunling Shen, and Jianwei Wang
Subjects: Saffold cardiovirus, viruses, pediatric patients, respiratory tract infections, dispatch, China, Medicine, Infectious and parasitic diseases, RC109-216
Abstract: To clarify the potential for respiratory transmission of Saffold cardiovirus (SAFV) and characterize the pathogen, we analyzed respiratory specimens from 1,558 pediatric patients in Beijing. We detected SAFV in 7 (0.5%) patients and identified lineages 1–3. However, because 3 patients had co-infections, we could not definitively say SAFV caused disease.
Published: 2010
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6. Diagnostic values of the QuantiFERON-TB Gold In-tube assay carried out in China for diagnosing pulmonary tuberculosis.

Author: Hui Xia, Xiaomen Wang, Fabin Li, Christophe Longuet, Guy Vernet, Delia Goletti, Yanlin Zhao, and Philippe H Lagrange
Subjects: Medicine, Science
Abstract: BACKGROUND:Interferon-release assays (IGRAs) for diagnosing active pulmonary tuberculosis (PTB) are not yet fully validated, particularly in high TB-endemic areas as the People's Republic of China (PRC). The aim of this report was to assess the performance of the QuantiFERON-TB Gold In-tube (QFT-GIT) and tuberculin skin test (TST), in addition to microbiological results, as contributors for diagnosing active PTB in the PRC. METHODS/PRINCIPAL FINDINGS:A total of 300 PTB patients, 41 disease controls (DC) and 59 healthy community controls (HCC) were included prospectively between May 2010 and April 2011 from two provinces of the PRC (Heilongjiang and Zhejiang). The QFT-GIT and TST yielded an overall sensitivity for active TB of 80.9% and 86.2%, and a specificity of 36.6% and 26.8%, respectively. The province of origin and smear microscopy status did not significantly impact the diagnostic values for PTB. However, using the TST with a 10 mm cut-off point, a significantly higher proportion of LTBI was observed in the DC than the HCC (p=0.01). Discordant results between the QFT-GIT and TST were found among 1/3 of the PTB, HCC and DC. Two-thirds of the individuals presented TST-positive/QFT-GIT-negative discordant results. The TST-negative/QFT-GIT-positive result was not associated with age or bacillary load. Cumulative QFT-GIT and TST positive results increased the overall sensitivity (95.9%), but it was associated with a dramatic decrease of the overall specificity (24.8%) leading to a suboptimal PPV (80.1%) and a low NPV (61.1%). CONCLUSIONS/SIGNIFICANCE:The usefulness of the QFT-GIT to diagnose active TB in high TB-endemic countries remains doubtful because like the TST, the QFT-GIT cannot distinguish between LTBI and active TB. Used as single stand-alone tests, both the QFT-GIT and TST have very limited roles in the diagnosis of active PTB. However, the combined use of SM, the TST and QFT-GIT may allow for the exclusion of ATB.
Published: 2015
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7. Human Enterovirus Genotype C104, China

Author: Zichun Xiang, Zhengde Xie, Zhong Wang, Lili Ren, Yan Xiao, Linlin Li, Guy Vernet, Gláucia Paranhos-Baccalà, Kunling Shen, and Jianwei Wang
Subjects: Picornaviridae, enterovirus, viruses, EV-C104, China, human enterovirus, Medicine, Infectious and parasitic diseases, RC109-216
Published: 2013
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8. Bocavirus in Children with Respiratory Tract Infections

Author: Li Guo, Richard Gonzalez, Zhengde Xie, Hongli Zhou, Chunyan Liu, Chao Wu, Gláucia Paranhos-Baccalà, Guy Vernet, Kunling Shen, Qi Jin, and Jianwei Wang
Subjects: viruses, respiratory infections, detection, bocavirus, human bocavirus, respiratory tract infections, Medicine, Infectious and parasitic diseases, RC109-216
Published: 2011
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9. Conserved B-cell epitopes among human bocavirus species indicate potential diagnostic targets.

Author: Zhuo Zhou, Xin Gao, Yaying Wang, Hongli Zhou, Chao Wu, Gláucia Paranhos-Baccalà, Guy Vernet, Li Guo, and Jianwei Wang
Subjects: Medicine, Science
Abstract: BACKGROUND: Human bocavirus species 1-4 (HBoV1-4) have been associated with respiratory and enteric infections in children. However, the immunological mechanisms in response to HBoV infections are not fully understood. Though previous studies have shown cross-reactivities between HBoV species, the epitopes responsible for this phenomenon remain unknown. In this study, we used genomic and immunologic approaches to identify the reactive epitopes conserved across multiple HBoV species and explored their potential as the basis of a novel diagnostic test for HBoVs. METHODOLOGY/PRINCIPAL FINDINGS: We generated HBoV1-3 VP2 gene fragment phage display libraries (GFPDLs) and used these libraries to analyze mouse antisera against VP2 protein of HBoV1, 2, and 3, and human sera positive for HBoVs. Using this approach, we mapped four epitope clusters of HBoVs and identified two immunodominant peptides--P1 (¹MSDTDIQDQQPDTVDAPQNT²⁰), and P2 (¹⁶²EHAYPNASHPWDEDVMPDL¹⁸⁰)--that are conserved among HBoV1-4. To confirm epitope immunogenicity, we immunized mice with the immunodominant P1 and P2 peptides identified in our screen and found that they elicited high titer antibodies in mice. These two antibodies could only recognize the VP2 of HBoV 1-4 in Western blot assays, rather than those of the two other parvoviruses human parvovirus B19 and human parvovirus 4 (PARV4). Based on our findings, we evaluated epitope-based peptide-IgM ELISAs as potential diagnostic tools for HBoVs IgM antibodies. We found that the P1+P2-IgM ELISA showed a higher sensitivity and specificity in HBoVs IgM detection than the assays using a single peptide. CONCLUSIONS/SIGNIFICANCE: The identification of the conserved B-cell epitopes among human bocavirus species contributes to our understanding of immunological cross-reactivities of HBoVs, and provides important insights for the development of HBoV diagnostic tools.
Published: 2014
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10. A toolbox for tuberculosis (TB) diagnosis: an Indian multi-centric study (2006-2008); evaluation of serological assays based on PGL-Tb1 and ESAT-6/CFP10 antigens for TB diagnosis.

Author: Philippe H Lagrange, Satheesh K Thangaraj, Rajeshwar Dayal, Alaka Deshpande, Nirmal K Ganguly, Enrico Girardi, Beenu Joshi, Kiran Katoch, Vishwa M Katoch, Manoj Kumar, Vemu Lakshmi, Marc Leportier, Christophe Longuet, Subbalaxmi V S Malladi, Deepali Mukerjee, Deepthi Nair, Alamelu Raja, Balambal Raman, Camilla Rodrigues, Pratibha Sharma, Amit Singh, Sarman Singh, Archana Sodha, Basirudeen Syed Ahamed Kabeer, Guy Vernet, and Delia Goletti
Subjects: Medicine, Science
Abstract: BACKGROUND: The aim of this multi-centric prospective study in India was to assess the accuracy of a serological test as an additional tool for diagnosing active tuberculosis (ATB). In particular, an assay based on ELISA using a phenolic glycolipid (PGL-Tb1) or a fusion protein (ESAT-6/CFP10) was compared to the tuberculin skin test (TST) and the microbiological results according to HIV status. METHODS: Individuals with and without ATB and HIV infection were enrolled. Serology and TST results were analyzed per se and in combination with the microbiological data. RESULTS: Among the 778 ATB patients, 102 were HIV-infected, 316 HIV-uninfected and 360 had an HIV-unknown status. Of the 945 non-ATB subjects, 559 were at low risk (community adults) and 386 at high risk of M. tuberculosis exposure. Among those with ATB, the sensitivity of ELISA-PGL-Tb1 for ATB was higher than that of ELISA-ESAT-6/CFP10, both in HIV-infected (72.3% versus 63.7%, p = 0.29) and HIV-uninfected/HIV-unknown groups (40.5% versus 28.6%; p
Published: 2014
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11. Saffold Cardiovirus in Children with Acute Gastroenteritis, Beijing, China

Author: Lili Ren, Richard Gonzalez, Yan Xiao, Xiwei Xu, Lan Chen, Guy Vernet, Gláucia Paranhos-Baccalà, Qi Jin, and Jianwei Wang
Subjects: Saffold cardiovirus, children, acute gastroenteritis, viruses, Beijing, China, Medicine, Infectious and parasitic diseases, RC109-216
Abstract: To understand Saffold cardiovirus (SAFV) distribution, prevalence, and clinical relevance in China, we retrospectively studied SAFV in children with acute gastroenteritis and found SAFV in 12 (3.2%) of 373. Sequence homology of virus protein 1 genes suggested these strains belong to the SAFV-1 sublineage. SAFVs were found in samples positive for other diarrhea-causing viruses.
Published: 2009
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12. Human Rhinovirus Group C Infection in Children with Lower Respiratory Tract Infection

Author: Zichun Xiang, Richard Gonzalez, Zhengde Xie, Yan Xiao, Lan Chen, Yongjun Li, Chunyan Liu, Yinghui Hu, Yuan Yao, Suyun Qian, Rong Geng, Guy Vernet, Gláucia Paranhos-Baccalà, Kunling Shen, Qi Jin, and Jianwei Wang
Subjects: HRV-C, rhinovirus, infection, children, lower respiratory tract, letter, Medicine, Infectious and parasitic diseases, RC109-216
Published: 2008
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13. A toolbox for tuberculosis (TB) diagnosis: an Indian multicentric study (2006-2008). Evaluation of QuantiFERON-TB gold in tube for TB diagnosis.

Author: Philippe H Lagrange, Satheesh K Thangaraj, Rajeshwar Dayal, Alaka Deshpande, Nirmal K Ganguly, Enrico Girardi, Beenu Joshi, Kiran Katoch, Vishwa M Katoch, Manoj Kumar, Vemu Lakshmi, Marc Leportier, Christophe Longuet, Subbalaxmi V S Malladi, Deepali Mukerjee, Deepthi Nair, Alamelu Raja, Balambal Raman, Camilla Rodrigues, Pratibha Sharma, Amit Singh, Sarman Singh, Archana Sodha, Basirudeen Syed Ahamed Kabeer, Guy Vernet, and Delia Goletti
Subjects: Medicine, Science
Abstract: BACKGROUND: The aim of this multicentric prospective study in India was to assess the performance of the QuantiFERON TB-Gold in tube (QFT-GIT), Tuberculin Skin Test (TST) and microbiological results as additional tools for diagnosing active tuberculosis (TB) and latent infection (LTBI) according to Human Immunodeficiency Virus (HIV) status. METHODS: Individuals with and without active TB and HIV infection were enrolled between 2006-2008. QFT-GIT and TST results were analyzed per se and in combination with microbiological data. RESULTS: Among the 276 individuals (96 active pulmonary TB and 180 no active TB) tested by QFT-GIT, 18 indeterminate results (6.5%) were found, more significantly numerous in the HIV-infected (15/92; 16.3%) than the HIV-uninfected (3/184; 1.6%)(p
Published: 2013
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14. Multiple genomic recombination events in the evolution of saffold cardiovirus.

Author: Lili Ren, Yan Xiao, Jianguo Li, Lan Chen, Jing Zhang, Guy Vernet, and Jianwei Wang
Subjects: Medicine, Science
Abstract: BACKGROUND: Saffold cardiovirus (SAFV) is a new human cardiovirus with 11 identified genotypes. Little is known about the natural history and pathogenicity of SAFVs. METHODOLOGY/PRINCIPAL FINDINGS: We sequenced the genome of five SAFV-1 strains which were identified from fecal samples taken from children with viral diarrhea in Beijing, China between March 2006 and November 2007, and analyzed the phylogenetic and phylodynamic properties of SAFVs using the genome sequences of every known SAFV genotypes. We identified multiple recombination events in our SAFV-1 strains, specifically recombination between SAFV-2, -3, -4, -9, -10 and the prototype SAFV-1 strain in the VP4 region and recombination between SAFV-4, -6, -8, -10, -11 and prototype SAFV-1 in the VP1/2A region. Notably, recombination in the structural gene VP4 is a rare event in Cardiovirus. The ratio of nonsynonymous substitutions to synonymous substitutions indicates a purifying selection of the SAFV genome. Phylogenetic and molecular clock analysis indicates the existence of at least two subclades of SAFV-1 with different origins. Subclade 1 includes two strains isolated from Pakistan, whereas subclade 2 includes the prototype strain and strains isolated in China, Pakistan, and Afghanistan. The most recent common ancestor of all SAFV genotypes dates to the 1710s, and SAFV-1, -2, and -3 to the 1940s, 1950s, and 1960s, respectively. No obvious relationship between variation and pathogenicity exists in the critical domains of the CD and EF loops of viral capsid proteins or the multi-functional proteins L based on amino acid sequence identity comparison between SAFV genotypes. CONCLUSIONS/SIGNIFICANCE: Our findings suggest that intertypic recombination plays an important role in the diversity of SAFVs, highlighting the diversity of the five strains with the previously described SAFV-1 strains.
Published: 2013
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15. Identification of a nonstructural DNA-binding protein (DBP) as an antigen with diagnostic potential for human adenovirus.

Author: Li Guo, Chengjun Wu, Hongli Zhou, Chao Wu, Gláucia Paranhos-Baccalà, Guy Vernet, Qi Jin, Jianwei Wang, and Tao Hung
Subjects: Medicine, Science
Abstract: BACKGROUND: Human adenoviruses (HAdVs) have been implicated as important agents in a wide range of human illnesses. To date, 58 distinct HAdV serotypes have been identified and can be grouped into six species. For the immunological diagnosis of adenoviruses, the hexon protein, a structural protein, has been used. The potential of other HAdV proteins has not been fully addressed. METHODOLOGY/PRINCIPAL FINDINGS: In this study, a nonstructural antigenic protein, the DNA binding protein (DBP) of human adenovirus 5 and 35 (Ad5, Ad35) - was identified using immunoproteomic technology. The expression of Ad5 and Ad35 DBP in insect cells could be detected by rhesus monkey serum antibodies and healthy adult human serum positive for Ad5 and Ad35. Recombinant DBPs elicited high titer antibodies in mice. Their conserved domain displayed immunological cross-reactions with heterologous DBP antibodies in Western blot assays. DBP-IgM ELISA showed higher sensitivity adenovirus IgM detection than the commercial Adenovirus IgM Human ELISA Kit. A Western blot method developed based on Ad5 DBP was highly consistent with (χ(2) = 44.9, P
Published: 2013
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16. Influenza A/H1N1 2009 pandemic and respiratory virus infections, Beijing, 2009-2010.

Author: Yaowu Yang, Zhong Wang, Lili Ren, Wei Wang, Guy Vernet, Gláucia Paranhos-Baccalà, Qi Jin, and Jianwei Wang
Subjects: Medicine, Science
Abstract: To determine the role of the pandemic influenza A/H1N1 2009 (A/H1N1 2009pdm) in acute respiratory tract infections (ARTIs) and its impact on the epidemic of seasonal influenza viruses and other common respiratory viruses, nasal and throat swabs taken from 7,776 patients with suspected viral ARTIs from 2006 through 2010 in Beijing, China were screened by real-time PCR for influenza virus typing and subtyping and by multiplex or single PCR tests for other common respiratory viruses. We observed a distinctive dual peak pattern of influenza epidemic during the A/H1N1 2009pdm in Beijing, China, which was formed by the A/H1N1 2009pdm, and a subsequent influenza B epidemic in year 2009/2010. Our analysis also shows a small peak formed by a seasonal H3N2 epidemic prior to the A/H1N1 2009pdm peak. Parallel detection of multiple respiratory viruses shows that the epidemic of common respiratory viruses, except human rhinovirus, was delayed during the pandemic of the A/H1N1 2009pdm. The H1N1 2009pdm mainly caused upper respiratory tract infections in the sampled patients; patients infected with H1N1 2009pdm had a higher percentage of cough than those infected with seasonal influenza or other respiratory viruses. Our findings indicate that A/H1N1 2009pdm and other respiratory viruses except human rhinovirus could interfere with each other during their transmission between human beings. Understanding the mechanisms and effects of such interference is needed for effective control of future influenza epidemics.
Published: 2012
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17. A toolbox for tuberculosis diagnosis: an Indian multicentric study (2006-2008): microbiological results.

Author: Philippe H Lagrange, Satheesh K Thangaraj, Rajeshwar Dayal, Alka Despande, Nirmal K Ganguly, Enrico Girardi, Beenu Joshi, Kiran Katoch, Vishwa M Katoch, Manoj Kumar, Vemu Lakshmi, Marc Leportier, Christophe Longuet, Subbalaxmi V S Malladi, Deepali Mukerjee, Deepthi Nair, Alamelu Raja, Balambal Raman, Camilla Rodrigues, Pratibha Sharma, Amit Singh, Sarman Singh, Archana Sodha, Basirudeen Syed Ahamed Kabeer, Guy Vernet, and Delia Goletti
Subjects: Medicine, Science
Abstract: BACKGROUND: The aim of this multicentric prospective study in India was to assess the value of several microbiological tools that contribute to the diagnosis of tuberculosis (TB) according to HIV status. METHODS: Standard microbiological tools on individual specimens were analyzed. RESULTS: Among the 807 patients with active TB, 131 were HIV-infected, 316 HIV-uninfected and 360 had HIV-unknown status. Among the 980 non-active TB subjects, 559 were at low risk and 421 were at high risk of M. tuberculosis (Mtb) exposure. Sensitivity of smear microscopy (SM) was significantly lower in HIV-infected (42.2%) than HIV-uninfected (75.9%) (p = 0.0001) and HIV-unknown pulmonary TB patients (61.4%) (p = 0.004). Specificity was 94.5% in non-TB patients and 100% in health care workers (HCW) and healthy family contacts. Automated liquid culture has significantly higher diagnostic performances than solid culture, measured by sensitivity (74.7% vs. 55.9%) (p = 0.0001) and shorter median time to detection (TTD) (12.0 vs. 34.0 days) (p = 0.0001). Specificity was 100% in HCW and cured-TB patients, but was lower in non-TB patients (89%) due to isolation of Mycobacteria other than tuberculosis (MOTT). TTD by both methods was related to AFB score. Contamination rate was low (1.4%). AccuProbe hybridization technique detected Mtb in almost all culture-positive specimens, but MOTT were found in 4.7% with a significantly higher frequency in HIV-infected (15%) than HIV-uninfected TB patients (0.5%) (p = 0.0007). Pre-test classification significantly increased the diagnostic value of all microbiological tests in pulmonary TB patients (p
Published: 2012
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18. Crimean-Congo hemorrhagic fever virus-infected hepatocytes induce ER-stress and apoptosis crosstalk.

Author: Raquel Rodrigues, Gláucia Paranhos-Baccalà, Guy Vernet, and Christophe N Peyrefitte
Subjects: Medicine, Science
Abstract: Crimean-Congo hemorrhagic fever virus (CCHFV) is a widely distributed tick-borne member of the Nairovirus genus (Bunyaviridae) with a high mortality rate in humans. CCHFV induces a severe disease in infected patients that includes, among other symptoms, massive liver necrosis and failure. The interaction between liver cells and CCHFV is therefore important for understanding the pathogenesis of this disease. Here, we described the in vitro CCHFV-infection and -replication in the hepatocyte cell line, Huh7, and the induced cellular and molecular response modulation. We found that CCHFV was able to infect and replicate to high titres and to induce a cytopathic effect (CPE). We also observed by flow cytometry and real time quantitative RT-PCR evidence of apoptosis, with the participation of the mitochondrial pathway. On the other hand, we showed that the replication of CCHFV in hepatocytes was able to interfere with the death receptor pathway of apoptosis. Furthermore, we found in CCHFV-infected cells the over-expression of PUMA, Noxa and CHOP suggesting the crosstalk between the ER-stress and mitochondrial apoptosis. By ELISA, we observed an increase of IL-8 in response to viral replication; however apoptosis was shown to be independent from IL-8 secretion. When we compared the induced cellular response between CCHFV and DUGV, a mild or non-pathogenic Nairovirus for humans, we found that the most striking difference was the absence of CPE and apoptosis. Despite the XBP1 splicing and PERK gene expression induced by DUGV, no ER-stress and apoptosis crosstalk was observed. Overall, these results suggest that CCHFV is able to induce ER-stress, activate inflammatory mediators and modulate both mitochondrial and death receptor pathways of apoptosis in hepatocyte cells, which may, in part, explain the role of the liver in the pathogenesis of CCHFV.
Published: 2012
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19. Differential seroprevalence of human bocavirus species 1-4 in Beijing, China.

Author: Li Guo, Yaying Wang, Hongli Zhou, Chao Wu, Jingdong Song, Jianguo Li, Gláucia Paranhos-Baccalà, Guy Vernet, Jianwei Wang, and Tao Hung
Subjects: Medicine, Science
Abstract: BACKGROUND: Four species of human bocaviruses (HBoV1-4) have been identified based on phylogenetic analysis since its first report in 2005. HBoV1 has been associated with respiratory disease, whereas HBoV2-4 are mainly detected in enteric infections. Although the prevalence of HBoVs in humans has been studied in some regions, it has not been well addressed globally. METHODOLOGY/PRINCIPAL FINDINGS: Cross-reactivity of anti-VP2 antibodies was detected between HBoV1, 2, 3, and 4 in mouse and human serum. The prevalence of specific anti-VP2 IgG antibodies against HBoV1-4 was determined in different age groups of healthy individuals aged 0-70 years old in Beijing, China, using a competition ELISA assay based on virus-like particles of HBoV1-4. The seroprevalence of HBoV1-4 was 50%, 36.9%, 28.7%, and 0.8%, respectively, in children aged 0-14 years (n = 244); whereas the seroprevalence of HBoV1-4 was 66.9%, 49.3%, 38.7% and 1.4%, respectively, in healthy adults (≥ 15 years old; n = 142). The seropositive rate of HBoV1 was higher than that of HBoV2, HBoV3, and HBoV4 in individuals older than 0.5 years. Furthermore, IgG seroconversion of HBoV1 (10/31, 32.3%), HBoV2 (8/31, 25.8%), and HBoV3 (2/31, 6.5%) was found in paired sera collected from children with respiratory tract infections who were positive for HBoV1 according to PCR analysis. CONCLUSIONS/SIGNIFICANCE: Our data indicate that HBoV1 is more prevalent than HBoV2, HBoV3, and HBoV4 in the population we sampled in Beijing, China, suggesting that HBoV species may play differential roles in disease.
Published: 2012
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20. Viral and atypical bacterial etiology of acute respiratory infections in children under 5 years old living in a rural tropical area of Madagascar.

Author: Jonathan Hoffmann, Henintsoa Rabezanahary, Martin Randriamarotia, Arsène Ratsimbasoa, Josette Najjar, Guy Vernet, Bénédicte Contamin, and Gláucia Paranhos-Baccalà
Subjects: Medicine, Science
Abstract: BACKGROUND: In Madagascar, very little is known about the etiology and prevalence of acute respiratory infections (ARIs) in a rural tropical area. Recent data are needed to determine the viral and atypical bacterial etiologies in children with defined clinical manifestations of ARIs. METHODS: During one year, we conducted a prospective study on ARIs in children between 2 to 59 months in the community hospital of Ampasimanjeva, located in the south-east of Madagascar. Respiratory samples were analyzed by multiplex real-time RT-PCR, including 18 viruses and 2 atypical bacteria. The various episodes of ARI were grouped into four clinical manifestations with well-documented diagnosis: "Community Acquired Pneumonia"(CAP, group I), "Other acute lower respiratory infections (Other ALRIs, group II)", "Upper respiratory tract infections with cough (URTIs with cough, group III)"and "Upper respiratory tract infections without cough (URTIs without cough, group IV)". RESULTS: 295 children were included in the study between February 2010 and February 2011. Viruses and/or atypical bacteria respiratory pathogens were detected in 74.6% of samples, the rate of co-infection was 27.3%. Human rhinovirus (HRV; 20.5%), metapneumovirus (HMPV A/B, 13.8%), coronaviruses (HCoV, 12.5%), parainfluenza virus (HPIV, 11.8%) and respiratory syncytial virus A and B (RSV A/B, 11.8%) were the most detected. HRV was predominantly single detected (23.8%) in all the clinical groups while HMPV A/B (23.9%) was mainly related to CAP (group I), HPIV (17.3%) to the "Other ALRIs" (group II), RSV A/B (19.5%) predominated in the group "URTIs with cough" (group III) and Adenovirus (HAdV, 17.8%) was mainly detected in the "without cough" (group IV). INTERPRETATION: This study describes for the first time the etiology of respiratory infections in febrile children under 5 years in a malaria rural area of Madagascar and highlights the role of respiratory viruses in a well clinically defined population of ARIs.
Published: 2012
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21. Evolutionary dynamics analysis of human metapneumovirus subtype A2: genetic evidence for its dominant epidemic.

Author: Jianguo Li, Lili Ren, Li Guo, Zichun Xiang, Gláucia Paranhos-Baccalà, Guy Vernet, and Jianwei Wang
Subjects: Medicine, Science
Abstract: Human metapneumovirus (hMPV) is a respiratory viral pathogen in children worldwide. hMPV is divided into four subtypes: hMPV_A1, hMPV_A2, hMPV_B1, and hMPV_B2. hMPV_A2 can be further divided into hMPV_A2a and A2b based on phylogenetic analysis. The typical prevalence pattern of hMPV involves a shift of the predominant subtype within one or two years. However, hMPV_A2, in particular hMPV_A2b, has circulated worldwide with a several years long term high epidemic. To study this distinct epidemic behavior of hMPV_A2, we analyzed 294 sequences of partial G genes of the virus from different countries. Molecular evolutionary data indicates that hMPV_A2 evolved toward heterogeneity faster than the other subtypes. Specifically, a bayesian skyline plot analysis revealed that hMPV_A2 has undergone a generally upward fluctuation since 1997, whereas the other subtypes experienced only one upward fluctuation. Although hMPV_A2 showed a lower value of mean dN/dS than the other subtypes, it had the largest number of positive selection sites. Meanwhile, various styles of mutation were observed in the mutation hotspots of hMPV_A2b. Bayesian phylogeography analysis also revealed two fusions of diffusion routes of hMPV_A2b in India (June 2006) and Beijing, China (June 2008). Sequences of hMPV_A2b retrieved from GenBank boosted simultaneously with the two fusions respectively, indicating that fusion of genetic transmission routes from different regions improved survival of hMPV_A2. Epidemic and evolutionary dynamics of hMPV_A2b were similar to those of hMPV_A2. Overall, our findings provide important molecular insights into hMPV epidemics and viral variation, and explain the occurrence of an atypical epidemic of hMPV_A2, particularly hMPV_A2b.
Published: 2012
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22. The human metapneumovirus matrix protein stimulates the inflammatory immune response in vitro.

Author: Audrey Bagnaud-Baule, Olivier Reynard, Magali Perret, Jean-Luc Berland, Mimoun Maache, Christophe Peyrefitte, Guy Vernet, Viktor Volchkov, and Gláucia Paranhos-Baccalà
Subjects: Medicine, Science
Abstract: Each year, during winter months, human Metapneumovirus (hMPV) is associated with epidemics of bronchiolitis resulting in the hospitalization of many infants. Bronchiolitis is an acute illness of the lower respiratory tract with a consequent inflammation of the bronchioles. The rapid onset of inflammation suggests the innate immune response may have a role to play in the pathogenesis of this hMPV infection. Since, the matrix protein is one of the most abundant proteins in the Paramyxoviridae family virion, we hypothesized that the inflammatory modulation observed in hMPV infected patients may be partly associated with the matrix protein (M-hMPV) response. By western blot analysis, we detected a soluble form of M-hMPV released from hMPV infected cell as well as from M-hMPV transfected HEK 293T cells suggesting that M-hMPV may be directly in contact with antigen presenting cells (APCs) during the course of infection. Moreover, flow cytometry and confocal microscopy allowed determining that M-hMPV was taken up by dendritic cells (moDCs) and macrophages inducing their activation. Furthermore, these moDCs enter into a maturation process inducing the secretion of a broad range of inflammatory cytokines when exposed to M-hMPV. Additionally, M-hMPV activated DCs were shown to stimulate IL-2 and IFN-γ production by allogeneic T lymphocytes. This M-hMPV-mediated activation and antigen presentation of APCs may in part explain the marked inflammatory immune response observed in pathology induced by hMPV in patients.
Published: 2011
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23. Identification of a highly conserved H1 subtype-specific epitope with diagnostic potential in the hemagglutinin protein of influenza A virus.

Author: Rongmao Zhao, Shujuan Cui, Li Guo, Chao Wu, Richard Gonzalez, Gláucia Paranhos-Baccalà, Guy Vernet, Jianwei Wang, and Tao Hung
Subjects: Medicine, Science
Abstract: Subtype specificity of influenza A virus (IAV) is determined by its two surface glycoproteins, hemagglutinin (HA) and neuraminidase (NA). For HA, 16 distinct subtypes (H1-H16) exist, while nine exist for NA. The epidemic strains of H1N1 IAV change frequently and cause annual seasonal epidemics as well as occasional pandemics, such as the notorious 1918 influenza pandemic. The recent introduction of pandemic A/H1N1 IAV (H1N1pdm virus) into humans re-emphasizes the public health concern about H1N1 IAV. Several studies have identified conserved epitopes within specific HA subtypes that can be used for diagnostics. However, immune specific epitopes in H1N1 IAV have not been completely assessed. In this study, linear epitopes on the H1N1pdm viral HA protein were identified by peptide scanning using libraries of overlapping peptides against convalescent sera from H1N1pdm patients. One epitope, P5 (aa 58-72) was found to be immunodominant in patients and to evoke high titer antibodies in mice. Multiple sequence alignments and in silico coverage analysis showed that this epitope is highly conserved in influenza H1 HA [with a coverage of 91.6% (9,860/10,767)] and almost completely absent in other subtypes [with a coverage of 3.3% (792/23,895)]. This previously unidentified linear epitope is located outside the five well-recognized antigenic sites in HA. A peptide ELISA method based on this epitope was developed and showed high correlation (χ(2) = 51.81, P
Published: 2011
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24. Is IP-10 an accurate marker for detecting M. tuberculosis-specific response in HIV-infected persons?

Author: Delia Goletti, Alamelu Raja, Basirudeen Syed Ahamed Kabeer, Camilla Rodrigues, Archana Sodha, Stefania Carrara, Guy Vernet, Christophe Longuet, Giuseppe Ippolito, Satheesh Thangaraj, Marc Leportier, Enrico Girardi, and Philippe Henri Lagrange
Subjects: Medicine, Science
Abstract: BackgroundThe suboptimal sensitivity of Interferon (IFN)-γ-based in-vitro assays, especially in immunocompromised individuals, emphasizes the need for alternative markers for diagnosing tuberculosis (TB). The objective of this study was to evaluate whether interferon-inducible protein (IP)-10, monocyte chemotactic protein (MCP)-2 and interleukin (IL)-2 can be useful biomarkers for evaluating a specific response to RD1 antigens associated to active TB disease in HIV-infected individuals.Methodology/principal findingsThe study was carried out in India, the country with the highest TB burden in the world. Sixty-six HIV-infected individuals were prospectively enrolled, 28 with active-pulmonary-TB and 38 without. The whole blood assay based on RD1-selected peptides (experimental test) and QuantiFERON-TB Gold In tube (QFT-IT) was performed. Plasma was harvested at day-1-post-culture and soluble factors were evaluated by ELISA. The results indicate that by detecting IP-10, the sensitivity of the experimental test and QFT-antigen (75% and 85.7% respectively) for active TB was higher compared to the same assays based on IFN-γ (42.9% and 60.7% respectively) and was not influenced by the ability to respond to the mitogen. By detecting IP-10, the specificity of the experimental test and QFT-antigen (57.9% and 13.2% respectively) for active TB was lower than what was reported for the same assays using IFN-γ-detection (78.9% and 68.4% respectively). On the other side, in vitro IL-2 and MCP-2 responses were not significantly associated with active TB.ConclusionsHIV infection does not impair RD1-specific response detected by IP-10, while it significantly decreases IFN-γ-mediated responses. At the moment it is unclear whether higher detection is related to higher sensitivity or lower specificity of the assay. Further studies in high and low TB endemic countries are needed to elucidate this.
Published: 2010
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25. Christophe Mérieux Conference, Trends in Virology

Author: Guy Vernet and Fabien Zoulim
Subjects: Emerging viral epidemics, surveillance, response, conference summary, Medicine, Infectious and parasitic diseases, RC109-216
Published: 2008
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26. WU Polyomavirus in Fecal Specimens of Children with Acute Gastroenteritis, China

Author: Lili Ren, Richard Gonzalez, Xiwei Xu, Jianguo Li, Jing Zhang, Guy Vernet, Gláucia Paranhos-Baccalà, Qi Jin, and Jianwei Wang
Subjects: WU polyomavirus, feces, fecal specimens, gastroenteritis, letter, China, Medicine, Infectious and parasitic diseases, RC109-216
Published: 2009
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27. Very high relative seroprevalence of anti-SARS-CoV-2 antibodies among communities in Bangui, Central African Republic

Author: Pr. Emmanuel Nakoune, Brice Martial Yambiyo, Alexandre Manirakiza, Joella Namsenei, Pr. Jean de Dieu Longo, Marie-Astrid Vernet, Jean Baptiste Roungou, Pr. Narcisse Patrice Komas, Modeste Bouhouda, Christian Malaka, Cathy Sandra Goimelle Coti-Reckoundji, Pr. Henri Saint-Calver Diemer, Guy Vernet, Pr. Gérard Grésenguet, and Marie Roseline Darnycka Belizaire
Subjects: History, medicine.medical_specialty, education.field_of_study, Polymers and Plastics, business.industry, Public health, Population, Industrial and Manufacturing Engineering, Herd immunity, Medicine, Seroprevalence, Residence, Cumulative incidence, Business and International Management, business, education, Serostatus, Sex ratio, Demography
Abstract: BackgroundLarge-scale population-based seroprevalence studies of SARS-CoV-2 are essential to characterize the cumulative incidence of SARS-CoV-2 infection and to extrapolate the prevalence of presumptive immunity at the population level.ObjectiveThe objective of our survey was to estimate the cumulative population immunity for COVID-19 and to identify individual characteristics associated with a positive serostatus.MethodThis was a clustered cross-sectional study conducted from July 12 to August 20, 2021, in households in the city of Bangui, the capital of the Central African Republic. Information regarding demographic characteristics (age, gender, and place of residence), comorbidities (chronic diseases) was collected. A venous blood sample was obtained for each participant to determine the level of total anti-SARS-CoV-2 antibodies using a WANTAI SARS-CoV-2 Ab ELISA kit.ResultsAll up, 799 participants were surveyed. The average age was 27 years, and 45·8% of the respondents were male (sex ratio: 0.8). The overall proportion of respondents with a positive serostatus was 74·1%. Participants over 20 years of age were twice as likely to have a positive serostatus, with an OR of 2.· ·2 (95% CI: [1·6, 3·1]).InterpretationThe results of this survey revealed a high cumulative level of immunity in Bangui, thus indicating a significant degree of spread of SARS-CoV-2 in the population. The public health implications of this high level of immunity to SARS-CoV-2, particularly on its variants burden, remain to be determined.FundingThis study was funded by the French Ministry for Europe and Foreign Affairs through the REPAIR COVID-19-Africa project coordinated by the Pasteur International Network association.
Published: 2021
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28. Precision Public Health Through Serological Biomarkers: An Integrated Surveillance Platform to Inform Public Health Interventions

Author: Barbara Jauregui, Benjamin F. Arnold, Guy Vernet, Isabel Rodriguez-Barraquer, Kathryn Banke, Robert H. Hall, Henrik Salje, Daniel T. Leung, Djibril Wade, Adriana Costero-Saint Denis, Kyla Hayford, Andrew S. Azman, and Kirsten E. Wiens
Subjects: medicine.medical_specialty, business.industry, Environmental health, Serological biomarkers, Public health, Public health interventions, other, Medicine, Sero epidemiology, business
Abstract: The use of biomarkers to measure immune responses in serum is crucial for understanding population-level exposure and susceptibility to human pathogens. Advances in sample collection, multiplex testing, and computational modeling are transforming serosurveillance into a powerful tool for public health program design and response to infectious threats. In July 2018, 70 scientists from 20 countries met to perform a landscape analysis of approaches that support an integrated serosurveillance platform, including the consideration of issues for successful implementation. Here, we summarize the group’s insights and proposed roadmap for implementation, including objectives, technical requirements, ethical issues, logistical considerations, and monitoring and evaluation.
Published: 2021
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29. Evaluation of the performances of twelve rapid diagnostic tests for diagnosis of HIV infection in Yaounde, Cameroon

Author: Laure Ngono, Marie-Astrid Vernet, Desirée Denise Mekinda-Gometi, Cyprien Kengne Nde, Guy Vernet, Richard Njouom, Patrice Tchendjou, and Serge Alain Sadeuh-Mba
Subjects: 0301 basic medicine, medicine.medical_specialty, Time Factors, Human immunodeficiency virus (HIV), HIV Infections, Hiv testing, Nucleic Acid Testing, medicine.disease_cause, Sensitivity and Specificity, Plasma, 03 medical and health sciences, 0302 clinical medicine, Predictive Value of Tests, Virology, Internal medicine, parasitic diseases, Humans, Mass Screening, Medicine, Cameroon, 030212 general & internal medicine, Plasma samples, Diagnostic Tests, Routine, business.industry, virus diseases, Diagnostic test, Gold standard (test), 030112 virology, Predictive value, Immunology, Enzyme immunoassays, business
Abstract: According to the WHO/UNAIDS recommendations, an acceptable HIV rapid diagnostic tests (RDTs) has to perform a sensitivity≥99% and a specificity≥98%. Given the constant release of new RDTs for HIV testing in the market and the high HIV genetic diversity in Cameroon, it is interesting to monitor their performances in that setting. A total of 240 HIV positive (including 219 HIV-1 M, 15 HIV-1 O, 1 HIV-1 N, 1 HIV-1 M/O recombinant and 4 HIV-2) and 240 HIV negative plasma samples were used to evaluate twelve routinely used RDTs in Cameroon. A reference algorithm combining Enzyme Immunoassays and nucleic acid testing was used as gold standard. The sensitivity, specificity, positive predictive value, and negative predictive value of the twelve RDTs evaluated varied between 93.7 and 100%; 95.8 and 100%; 96.0 and 100%, and 94.1 and 100%, respectively. Five out of the twelve RDTs could not detect some HIV-1 O variants, one of them failed to detect an HIV-2 variant while all them efficiently detected HIV-1 N and HIV M/O recombinant. Our findings underscore the need to monitor the performances of RDTs to be used for HIV testing in Cameroon using locally obtained well-characterized samples panels.
Published: 2017
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30. Importation and outbreak of wild polioviruses from 2000 to 2014 and interruption of transmission in Cameroon

Author: Richard Njouom, Cara C. Burns, Marcellin Nimpa Mengouo, Marie Claire Endegue-Zanga, Guy Vernet, François-Xavier Etoa, Jane Iber, Serge Alain Sadeuh-Mba, Francis Delpeyroux, Marycelin Baba, Maurice Demanou, Nicksy Gumede Moeletsi, and David Bukbuk
Subjects: Male, 0301 basic medicine, Adolescent, Genotype, Biology, medicine.disease_cause, Virus, Disease Outbreaks, law.invention, Feces, 03 medical and health sciences, 0302 clinical medicine, law, Virology, Disease Transmission, Infectious, medicine, Humans, Cameroon, 030212 general & internal medicine, Child, Poliovirus type, Retrospective Studies, Travel, Incidence, Incidence (epidemiology), Poliovirus, Infant, Newborn, Infant, Outbreak, Sequence Analysis, DNA, medicine.disease, Poliomyelitis, 030104 developmental biology, Infectious Diseases, Transmission (mechanics), Immunization, Child, Preschool, Female, Topography, Medical
Abstract: Background Efficient implementation of the global eradication strategies consisting of Acute Flaccid Paralysis (AFP) surveillance and mass immunization campaigns led to interruption of indigenous wild poliovirus transmission in Cameroon in 1999. Objectives This study describes type 1 and type 3 wild poliovirus (WPV) importation, incidence, geographic distribution and control since the original interruption of transmission in Cameroon. Study design Stool samples from AFP patients under the age of 15 years in Cameroon were collected nationwide and subjected to virus isolation on RD and L20B cell cultures. Resulting virus isolates were typed by intratypic differentiation (ITD) and analysis of the VP1 coding sequence of the viral genome. Surveillance data originating from Cameroon between 2000 and 2014 were considered for retrospective descriptive analyses. Results From 2003 to 2009, multiple WPV importation events from neighboring countries affected mainly in the northern regions of Cameroon but did not led to sustained local transmission. Throughout this period, 16 WPV1 and 5 WPV3 were detected and identified as members of multiple clusters within type-specific West Africa B genotypes (WEAF-B). In 2013–2014, a polio outbreak associated to a highly evolved (“orphan”) WPV1 affected four southern regions of Cameroon. Conclusions The appearance of highly evolved lineage of type 1 WPV suggests potential surveillance gap and underscore the need to maintain comprehensive polio immunization activities and sensitive surveillance systems in place as long as any country in the world remains endemic for WPV.
Published: 2016
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31. Absence of hybridization with the wild-type and mutant rpoB probes in the Genotype MTBDRplus assay detects 'disputed' rifampicin mutations

Author: Diane Wallace Taylor, Guy Vernet, Wilfred Fon Mbacham, Rose G. F. Leke, Ngu Njei Abanda, Sara Irene Eyangoh, Josiane Yvonne Djieugoué, Veronique Mbeng Penlap, Vedbar S. Khadka, Youping Deng, and Eric Walter Pefura-Yone
Subjects: 0301 basic medicine, Microbiology (medical), DNA, Bacterial, Genotype, 030106 microbiology, Mutant, Antitubercular Agents, Microbial Sensitivity Tests, medicine.disease_cause, Article, Mycobacterium tuberculosis, Diagnosis, Differential, 03 medical and health sciences, Bacterial Proteins, Drug Resistance, Bacterial, Tuberculosis, Multidrug-Resistant, medicine, polycyclic compounds, Isoniazid, Humans, Genetics, Mutation, Microbial Viability, biology, Base Sequence, Diagnostic Tests, Routine, Wild type, General Medicine, DNA-Directed RNA Polymerases, biology.organism_classification, rpoB, bacterial infections and mycoses, Phenotype, Infectious Diseases, Genes, Bacterial, Rifampin, Rifampicin, medicine.drug
Abstract: Objective Mycobacterium tuberculosis isolates that fail to hybridize to at least one rpoB wild-type or any mutation probe on the Genotype MTBDRplus strip are assumed to be rifampicin-resistant. However, the precise mutation(s) are unknown. We sought to identify the mutations in isolates with such hybridization patterns and determine if the mutations are associated with resistance to rifampicin. Methods In this study, 275 M. tuberculosis isolates were screened with the Genotype MTBDRplus assay to identify isolates with the hybridization pattern. These isolates were sequenced and their minimum inhibitory concentrations (MIC) determined using the Bactec MGIT 960 system. Results Among the 275 isolates tested, 15 (6%) isolates with the hybridization pattern were identified. Sequencing showed that failure to hybridize to rpoB wild-type probes resulted from the presence of ‘disputed' rifampicin mutations, which are mutations not always associated with a rifampicin-resistant phenotype. All, except 3/15, isolates had a rifampicin-resistant phenotype (MIC > 1 μg/mL). One of the three isolates with a rifampicin-susceptible phenotype had the same mutation at position 526 (His526Leu) as another isolate that had a rifampicin-resistant phenotype. Conclusion The recommendation of the Genotype MTBDRplus assay to assume rifampicin resistance based solely on failure to hybridize to rpoB wild-type probe allows the identification of important RIF-resistant isolates. About 20% (3/15) of such isolates could be missed by relying only on the standard MGIT 960 DST assay for drug susceptibility testing.
Published: 2017

32. HCV Ag quantification as a one-step procedure in diagnosing chronic hepatitis C infection in Cameroon: the ANRS 12336 study

Author: Frédéric Lissock, Léa Duchesne, Alexandre Soulier, Karine Lacombe, Pierre-Marie Girard, Gishlaine Flore Tamko-Mella, Sandrine Rallier, Lila Poiteau, Richard Njouom, Stéphane Chevaliez, Jean-Michel Pawlotsky, Guy Vernet, Nicolas Rouveau, Institut Pierre Louis d'Epidémiologie et de Santé Publique (iPLESP), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre Pasteur du Cameroun, Réseau International des Instituts Pasteur (RIIP), Service de bactériologie, virologie, hygiène [Mondor], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Dpt Research in resource-limited settings, ANRS France Recherche Nord & sud Sida-hiv hépatites, Services des Maladies Infectieuses et Tropicales [CHU Saint-Antoine], CHU Saint-Antoine [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), and HAL UPMC, Gestionnaire
Subjects: Male, diagnosis, Human immunodeficiency virus (HIV), HIV Infections, medicine.disease_cause, Gastroenterology, 0302 clinical medicine, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, 030212 general & internal medicine, Cameroon, HCV core antigen, biology, Sub-Saharan Africa, Coinfection, virus diseases, Middle Aged, Hepatitis B, 3. Good health, Infectious Diseases, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, 030211 gastroenterology & hepatology, Female, Hcv core antigen, Antibody, Research Article, Adult, medicine.medical_specialty, Context (language use), Sensitivity and Specificity, 03 medical and health sciences, co-infection, Chronic hepatitis, Internal medicine, medicine, chronic hepatitis C, Humans, Aged, Hepatitis B virus, business.industry, Public Health, Environmental and Occupational Health, HIV, [SDV.MHEP.HEG]Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology, Gold standard (test), Hepatitis C Antibodies, Hepatitis C, Chronic, [SDV.MHEP.HEG] Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology, digestive system diseases, Cross-Sectional Studies, ROC Curve, [SDV.SPEE] Life Sciences [q-bio]/Santé publique et épidémiologie, Concomitant, Immunology, biology.protein, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, Other, Hepatitis C Antigens, business, hepatitis B virus
Abstract: International audience; Introduction: The diagnostic procedure for chronic hepatitis C infection (CHC) usually combines anti-HCV antibody (HCV-Ab) and HCV-RNA measurement. Quantifying HCV core antigen (cAg) as a one-step procedure could shorten the diagnostic process. We aimed to assess the performance of cAg quantification in diagnosing CHC and how it is influenced by concomitant HIV or HBV infections. Methods: The cAg was quantified by an automated assay (Abbott Diagnostics) in 465 HCV-Ab negative serum samples and 544 HCV-RNA positive serum samples (n = 1009) collected in patients from the Pasteur Center in Cameroon, some of whom were infected by HBV or HIV. Its performance was evaluated in comparison to the gold standard (ELISA or PCR) by estimating its sensitivity (Se) and specificity (Sp), and by comparing the area under ROC (AUROC) curves in each patient population: HCV mono-infected, HCV-HBV and HIV-HCV co-infected. Results: Among the 465 HCV-Ab negative patients, 51 and 79 were HIV-and HBV-infected, respectively, whereas among the 544 patients with CHC, 27 and 28 were HIV-and HBV-infected, respectively. The Spearman ρ correlation coefficient between cAg and HCV-RNA was 0.75 (p < 0.00001). The assay had a sensitivity of 95.7% (95% CI: 93.2–97.5) and a specificity of 99.7% (95% CI: 98.1–10) in diagnosing CHC, corresponding to an AUROC of 0.99 (95% CI: 0.98–1.0). Being HIV-or HBV-infected did not impact the performance of cAg (Se = 96.4%, Sp = 96.2% and AUROC = 0.98 (95% CI: 0.95–1.0) in the HBV group, Se = 100%, Sp = 88.2% and AUROC = 0.99 (95% CI: 0.97–1.0) in the HIV group, p between AUROC = 0.69). Conclusions: The cAg quantification displayed a high specificity and sensitivity for the diagnosis of CHC in Cameroon, and its performance was not significantly modified by a concomitant HIV or HBV infection. In the context of CHC elimination on a global scale, using cAg quantification as a screening tool to directly identify CHC could be a reliable tool in a " test and treat " strategy.
Published: 2017
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33. Diagnostic accuracy and usefulness of the Genotype MTBDRplus assay in diagnosing multidrug-resistant tuberculosis in Cameroon? a cross-sectional study

Author: Sara Irene Eyangoh, Guy Vernet, Eric Walter Pefura-Yone, Wilfred Fon Mbacham, Ngu Njei Abanda, Eunjung Lim, Josiane Yvonne Djieugoué, Diane Wallace Taylor, Rose G. F. Leke, and Veronique Mbeng Penlap
Subjects: 0301 basic medicine, Male, Genotyping Techniques, Antitubercular Agents, rpoB mutations, Drug resistance, Mutation Rate, Genotype, Tuberculosis, Multidrug-Resistant, Cameroon, Cameroon. InhA promoter mutation, biology, INHA, Isoniazid, Pulmonary tuberculosis, Middle Aged, Multidrug-resistant tuberculosis (MDR-TB), 3. Good health, Infectious Diseases, Molecular Diagnostic Techniques, Female, medicine.symptom, Rifampin, Oxidoreductases, medicine.drug, Research Article, Adult, Tuberculosis, 030106 microbiology, Microbial Sensitivity Tests, Sensitivity and Specificity, lcsh:Infectious and parasitic diseases, Mycobacterium tuberculosis, 03 medical and health sciences, Bacterial Proteins, medicine, Humans, lcsh:RC109-216, Tuberculosis, Pulmonary, business.industry, Genotype MTBDRplus assay, biology.organism_classification, medicine.disease, bacterial infections and mycoses, Virology, KatG codon 315 mutation, Cross-Sectional Studies, Mutation, Sputum, business, Rifampicin
Abstract: Background Drug-resistant tuberculosis, especially multidrug-resistant tuberculosis (MDR-TB), is a major public health problem. Effective management of MDR-TB relies on accurate and rapid diagnosis. In this study, we assessed the diagnostic accuracy of the Genotype MTBDRplus assay in diagnosing MDR-TB in Cameroon, and then discuss on its utility within the diagnostic algorithm for MDR-TB. Methods In this cross-sectional study, 225 isolates of Mycobacterium tuberculosis cultured from sputum samples collected from new and previously treated pulmonary tuberculosis patients in Cameroon were used to determine the accuracy of the Genotype MTBDRplus assay. We compared the results of the Genotype MTBDRplus assay with those from the automated liquid culture BACTEC MGIT 960 SIRE system for sensitivity, specificity, and degree of agreement. The pattern of mutations associated with resistance to RIF and INH were also analyzed. Results The Genotype MTBDRplus assay correctly identified Rifampicin (RIF) resistance in 48/49 isolates (sensitivity, 98% [CI, 89%–100%]), Isoniazid (INH) resistance in 55/60 isolates (sensitivity 92% [CI, 82%–96%]), and MDR-TB in 46/49 (sensitivity, 94% [CI, 83%–98%]). The specificity for the detection of RIF-resistant and MDR-TB cases was 100% (CI, 98%–100%), while that of INH resistance was 99% (CI, 97%–100%). The agreement between the two tests for the detection of MDR-TB was very good (Kappa = 0.96 [CI, 0.92–1.00]). Among the 3 missed MDR-TB cases, the Genotype MTBDRplus assay classified two samples as RIF-monoresistant and one as INH monoresistant. The most frequent mutations detected by the Genotype MTBDRplus assay was the rpoB S531 L MUT3 41/49 (84%) in RIF-resistant isolates, and the KatG S315 T1 (MUT1) 35/55 (64%) and inhA C15T (MUT1) 20/55 (36%) mutations in INH-resistant isolates. Conclusion The Genotype MTBDRplus assay had good accuracy and could be used for the diagnosis of MDR-TB in Cameroon. For routine MDR-TB diagnosis, this assay could be used for Mycobacterium tuberculosis cultures containing contaminants, to complement culture-based drug susceptibility testing or to determine drug resistant mutations.
Published: 2017

34. Surveillance for Antimicrobial Drug Resistance in Under-Resourced Countries

Author: Catherine Mary, Ogobara K. Doumbo, Keith P. Klugman, Susan C. Morpeth, Zulfiqar A Bhutta, Guy Vernet, and Dany M. Altmann
Subjects: Microbiology (medical), Tuberculosis, Standardization, Epidemiology, lcsh:Medicine, Developing country, ComputerApplications_COMPUTERSINOTHERSYSTEMS, Resistance (psychoanalysis), Global Health, Communicable Diseases, lcsh:Infectious and parasitic diseases, 03 medical and health sciences, 0302 clinical medicine, Antibiotic resistance, under-resourced countries, Environmental health, diagnostics, Global health, Humans, Medicine, lcsh:RC109-216, antimicrobial resistance, 030212 general & internal medicine, Road map, Developing Countries, 0303 health sciences, 030306 microbiology, business.industry, lcsh:R, TheoryofComputation_GENERAL, Drug Resistance, Microbial, medicine.disease, Policy Review, 3. Good health, Infectious Diseases, Population Surveillance, surveillance, business, Malaria
Abstract: TOC summary: New programs can be improved by drawing on lessons from previous successful efforts., Antimicrobial drug resistance is usually not monitored in under-resourced countries because they lack surveillance networks, laboratory capacity, and appropriate diagnostics. This accelerating problem accounts for substantial number of excess deaths, especially among infants. Infections particularly affected by antimicrobial drug resistance include tuberculosis, malaria, severe acute respiratory infections, and sepsis caused by gram-negative bacteria. Nonetheless, mapping antimicrobial drug resistance is feasible in under-resourced countries, and lessons can be learned from previous successful efforts. Specimen shipping conditions, data standardization, absence of contamination, and adequate diagnostics must be ensured. As a first step toward solving this problem, we propose that a road map be created at the international level to strengthen antimicrobial resistance surveillance in under-resourced countries. This effort should include a research agenda; a map of existing networks and recommendations to unite them; and a communication plan for national, regional, and international organizations and funding agencies.
Published: 2014
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35. Enhancing research capacities in infectious diseases: The GABRIEL network, a joint approach to major local health issues in developing countries

Author: Graciela Russomando, Guy Vernet, Florence Komurian-Pradel, Dolla Karam Sarkis, Monidarin Chou, Jianwei Wang, Gláucia Paranhos-Baccalà, Wilfred Fon Mbacham, Souleymane Diallo, Pagbajab Nymadawa, Nina Grundmann, Jean W. Pape, Phimpha Paboriboune, Luc Hervé Samison, and Marilda M. Siqueira
Subjects: Microbiology (medical), Operations research, Epidemiology, business.industry, Public Health, Environmental and Occupational Health, Developing country, Public relations, Politics, Infectious Diseases, Research capacity, Sustainability, Medicine, Joint (building), business, Inefficiency
Abstract: Low income countries seldom possess adequate means to effectively carry out rigorous epidemiological studies in the fight against infectious diseases such as Tuberculosis and Acute Lower Respiratory Infections. To do so, they require solid health infrastructures and qualified researchers trained in the latest laboratory techniques. Where international collaborative programs presently operate in developing countries, the sustainability of research practices often represents a major challenge. The lack of coordination and support between political decision makers and scientific experts also accounts for the inefficiency of existing programs. The GABRIEL network was therefore recently created by the Fondation Merieux to help developing countries improve their research capacity for the detection, expertise, characterization, and surveillance of pathogens. The members of GABRIEL, being institutionally and geographically diverse, comprise laboratories, such as academic research laboratories, governmental institutions, and NGOs. GABRIEL's mission is to enhance health research capacity in developing countries by implementing sustainable research projects.
Published: 2013
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36. Chronic hepatitis B liver disease in patients living in the Amazon region: S gene mutations and genotypes characterization

Author: L. D. de Figueiredo Nicolete, Alcione de Oliveira dos Santos, Gláucia Paranhos-Baccalà, Carina Picelli, Marie Gauthier, Eduardo Rezende Honda, Guy Vernet, Deusilene Souza Vieira, and Juan Miguel Villalobos Salcedo
Subjects: Hepatitis B virus, Mutation, Molecular epidemiology, business.industry, Gene mutation, medicine.disease_cause, medicine.disease, Virology, Virus, Serology, Liver disease, Genotype, medicine, business
Abstract: The Amazon region is considered to be a high endemic area for Hepatitis B Virus (HBV) infections, Rondonia state having the highest prevalence. The aim of this study was to identify molecular genotypes and mutations in the S gene region of HBV viral genomes from 20 patients using a DNA microarray. Results: Serological tests showed that 88% of patients were HBeAg negative, 82% had anti-HBe antibodies and 33% were co-infected with Hepatitis Delta Virus. Sixteen percent of the patients were considered cirrhotic, and 11% have been transfused. The microarray technique identified the genotypes A (4 patients), D (7 patients) and F (7 patients) in 18 samples. Mutations were detected in all 3 genotypes and, overall, A159G, which has been associated with a reduced antigenicity of the virus, was detected most frequently. In genotype A, G119E was the most frequently detected mutation followed by mutations A159G, F134Y, W172C, Y161F and T143S. A159G was detected in all genotype D and F samples followed by mutations T143S, Y161F, N131T, T114S and G119E in genotype D and mutations T143S, Y161F, N131T, T114S and G119E in genotype F. Conclusion: The analysis of mutations repartition among genotypes suggests that some of them are preferentially or exclusively associated with genotype A, D or F. This type of tool is adapted for clinical and therapy monitoring of patient as well as for molecular epidemiology research on HBV.
Published: 2013
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37. Clinical features and prognostic factors of listeriosis: the MONALISA national prospective cohort study

Author: Odile Fremin-Batteux, Juliette Clarissou-Philippe, Benoît Jauhlac, Severine Guyetand, Jacques Gasnault, Corinne Haioun, Liamine Aissaoui, Marie-Christine Pages, Marie-Pierre Fos, Christian Rose, Didier Hubert, Marie-Rose Rothe, N. Bouziges, Benoît Huc, François Devianne, Sabine Bidart, Anne Forest, Kevin Bertrand, Mohamed Eldeghedy, Annick Verhaeghe, Caroline Malderet, Anne Bertrou, Bernard Guerquin, Catherine Duche, Muriel Archambaud, Rabea Cotteret, Olivier Toullalan, Yves Devaux, Smail Bergheul, Valérie Sivadon-Tardy, Pierre-Gilles Merville, Geneviève Blanchard-Marche, Didier Raoult, Bernard Hory, Florence Richardin, Evelyne Belle, Mohamed Menouar, K Guitteaud, Mohamad Mohty, Ambroise Montcriol, Max Laurin, Aurélia Picard, Jean-Paul Mira, Marie-Charlotte Chopin, Richard Bonnet, Michel Wolff, Sébastien Maillez, Jeanne Maugein, Véronique Leblond, Nicola Walid, Bernard Gauche, Mathieu Evillard, Hassen Jeddi, Anne Bourlet, Isabelle Grawey, Thierry Jault, Sandrine Hiret, Valerie Gaborieau, Véronique Boin-Gay, An Kim, Thierry Constans, Jean-François Gaide, Martine Giraud, Eric Meaudre Desgouttes, Alain Fur, Abdallah Maakaroun, Olivier Matray, Bertrand Maubert, Frédérique Péchinot, Aurelie Garbi, Claire Delbrouck, Benoît Grandclerc, Vincent Cadiergue, Hervé Lécuyer, Bernadette Grignon, Thierry Bensaid, Nicole Constantin, Yannick Chevalier, Hassène Rahmani, Thierry Levent, Joelle Desliers, Florence Van de Velde, Xavier Adhoute, Clara Andriau, Christophe Charasse, Rémi Vatan, Benoît Martha, Alain Lecis, Didier Albert, Romain Jacobs, Hélène Lefranc, Christian Martin, Nasseur Rezgui, Bertrand Pigeon, Catherine Le Henaff, Dominique Cassignard, Françoise Cotes, Eric Pujade Lauraine, Jean-François Gattault, Nicole Ferreira-Maident, Noémie Jourde-Chiche, Hélène Garrec, Olivier Darchen, Carole Schwebel, Marie-Christine Bezian, Patrick Daoud, Tsouria Becaid, Simone Laluque, David Broche, Christine Boisselier, Pascale Martres, Sarah Hammami, Brigitte Olivier, Jean-Marie Nkunzimana, Eric Monlun, Isabelle Marterl-Lafay, Marion Carboni, Marie-Françoise Mattei, Sandrine Castelin, Isabelle Barillot, Marie-Noelle Cufi, Thomas Kaiser, Catherine Herry, Pascal Hutin, Jean-Pierre Bronowicki, Bernard Branger, Pierre Thomas, Elie Zagdoun, Anne Goquelin, Ziad Assaf, Ingrid Croquet, Bruno Pozzetto, Thomas Similowski, Anne-Isabelle Briere, Marie-Thérèse Albertini, Mariam Blaka, Christelle Tassot, Anne Gaschet, Jean-Philippe Lavigne, Antoine Pujol, Philippe Colombat, Edouard Devaud, Hana Talabani-Boizot, François Barière, Anne-Marie Cordier, Philippe Gueudet, Georges Simon, Anne-Sophie Lipovac, Françoise Bandaly, Anne Beauplet-Lepage, Sylvie Prince, Charlotte Jouzel, Jean-Luc Deboutin, Patrick Zavadil, Louis Puybasset, Marie-Cécile Petit, Loïc Guillevin, Kamel Touati, Christophe Ntalu Nkato, Sylvie Carette, Jacques Vaucel, Chantal Delasalle, Marine Gross Goupil, Laurent Gutmann, Christiane Payen, Annick Barboteau, Firouzé Bani-Sadr, Christophe Legendre, Philippe Roulier, Elie Azria, Ibrahim Farah, Isabelle Rouquette-Vincent, Anne-Sophie Erena-Penet, Philippe Labadie, Eric Josien, Aicha Derragui, Mathieu Legrand, Odile Beyne-Rauzy, Jean-Marc Nabholtz, Marie-Joelle Demarcq, Olivier Garosi, Michel Deiber, Fabrice Chaix, Bertrand Souweine, Anne Collignon, Gisèle Renard, Mickael Jego, Gilles Bernardin, Anne Allart, Jocelyn Barrier, Marc Vasse, Philippe Ménager, Marc Wurmser, Abderkader Ouazir, Olivier Gontieron, Yvon Berland, Sébastien Trouiller, David Leysenne, Christophe Ozanon, Fanny Autret, Tahar Saghi, Loïc Dopeux, Sophie Benoit-Coustou, T. Fraisse, Christine Maillard, Karine Nikodijevic, Georges Kaltenbach, Angéline Jamet, Philippe Aucher, Julie Bottero, Marie-Claude Piffaut, Marianne Besnard, Florence Courillon, Marie Bonfils, Christine Ghevaert, Marie Destors, Eliette Jeanmaire, Franck Zerbib, Manuel-Luis Gameiro, T Prazuck, Laurent Mandin, Olivier Guisset, Marguerite Fines, Toufik Feddal, Agnès Jouffret, Louis Mesnard, Thomas Bourrée, Hasinrina Razafimahefa, Sylvestre Tigaud, Vincent Estève, Philippe Malherbe, Jean-Michel Salord, Pascal Adam, Bertrand Rozec, Michel Fuillet, Olivier Lemenand, Denis Quinsat, Ana Danalaché, Véronique Vialette, François Brosset, Patrick Messner Pellenc, Nicolas Heisel, Edouard Girard, Régine Martin, Olivier Garesslin, Catherine Mille, Alexandre Gascon, Marc Nicolino, Laurence Mouly, Claire Fabre, Bénédicte Ponceau, Marie-Etiennette Emeriau, Pascal Cathebras, Bérangère Bernardaud, Michèle Pérouse de Montclos, O. Arsene, Karine Grenet, Yazdan Yazdanpanah, Sten De Witte, Anne Scemla, Laurence Bouillet, Christophe Burucoa, Vincent Loffeier, Séverine Visentin, Luc Desfrere, Miloud Arabi, Frédérique Costa, Sylvie Lechat, Ali Chekroun, Raymond Ruimy, Marie, Jérôme Bizet, Xavier Nassif, Baihas Dib, Patrick Bert-Marcaz, Laurent Martin Lefèvre, Nicholas Sedillot, Blandine Cattier, Emilie Boidin, Daniel Sondag, Aude Bourrouillou, Alain Noirot, Franck Desemerie, Fréderic Heluwaert, Catherine Tamalet, Marc G. Berger, Jean-Daniel Lelièvre, Dominique Perotin, Abdelkader Bemrah, Alain Lozniewski, Bernard Borstein, Hanna Eid, Diana Suatean, Virginie Mignaut, Jean-Claude N'guyen, Valérie Le Goff, Laurent Teillet, Christophe Rolland, Gwenaël Alfonsi, Florence Lachenal, Philippe Bossi, Yves Botreau, Florence Doucet Populaire, Henry Jardel, Nicolas Gallo, Elias Jabre-Sikias, Michel Dupon, Hélène Brihier, Isabelle Patry, Alexandre Leclercq, Bernadette Tourrand, Christophe Roussel, Jean-Emmanuel Kurtz, Bénédicte Paindaveine, Simon Elhadad, Richard Sanchez, Eric Sgro, Pierre Berger, Valérie Murbach, Anne Holstein, Florence Martin, Taoufik Merabet, Amélie Benbara, Milagros Ferreyra, Laure Esposito, Pierre Delobel, Antoine Andremont, Marc Bourlière, Carole-Anne Boudy, Jean-Baptiste Gaulthier, Laurent Tacchini, Olivier Marpeau, Sonia Tesseydre, Marie-Pierre Coulhon, Nathalie Hodee, Marie-Chrsitine Conroy, Pierre Weinbreck, Roland Leclercq, Laurent Souply, Christian Bidault, Annie Elbez, Marie-Annick Lebreton, Patrick Brisou, Agnès Ferroni, Jean-Louis Pourriat, Nadia Anguel, Christian Noel, Philippe Jouvencel, Eric Pichard, Xavier Martin, Mathieu Coste, Pierre Zuber, Catherine Neuwirth, Jean-Pierre Hacot, Paul Aye, Jérôme Guinard, Yves Pean, Jean-Christophe Dengo, Fabrice Petassou, Didier Viole, Thierry Messiaen, Jean Beytout, Philippe Petitjean, Ferdinand Savare, Patrice Cuvillier, Sophie Coignard, Hélène Anglaret, Nassim Kamar, Elisabeth Chachaty, Karine Guimard, Louis Braem, Hacene Fezoui, Pierre Martin, Jean-Paul Viard, Claire Larroche, Nicolai Claudiu-Plesa, Thierry Benoit-Cattin, Olivier Moquet, Thierry Pasdeloup, David Rosay, Rodolphe Jean, Jean-Bernard Mariette, Marc Debouverie, Hervé Peltier, Mustapha Terki, Jacques Daleas, Valérie Dattin-Dorrière, Michel Vergnaud, Emmanuel Grimprel, Sylvène Rosselli, Jean-Marc Didier, Pierre Faurie, Luc Frimat, Aziza Mandjee, Sabine Etchemendy, Pierre Tissières, Jean Nakhleh, Sylvie Mariette, Christian Perronne, Bruno Carbonne, Nathalie Houssiaux-Maisonneuve, Tristan Ferry, S Beague, Anthony Sebban, Marie-Thérèse Hili, Jean-Michel De Kermadec, Lucien Brasme, Gilles Blaison, Caroline Garandeau, Jean-Pierre Sollet, Laurent Tronchon, Thierry Samson, Julien Gesquière, Nicolas Ettahar, Alain Créange, Etienne Laurens, Véronique Equy, Fréderic Bart, Bernard Bouffandeau, Christine Vaillant, Valerie Pesque, Jean-Marc Lalot, Marc Levy, Michel Kaidomar, Mihaela Saplacan, Sterenn Yvenou, Marie-Isabelle Steibach, Emmanuelle Cambau, Agnès Riche, François Fourrier, François Raffi, Mélissa Lalu, Henri Bérard, Danielle Clave, Jean-Claude Mouries, Martine Porcheron, Jean Cabalion, Richard Lamarca, Nathalie Canu, Jean-Baptiste Roseau, Annabelle Stoclin, Luca Luminitan Elena Lupean, Rémi Gebeile, Celia Salanoubat, Carole Marmouset, Pierre Bigot, Anne-Laure Breton, Pierre Kalfon, Colette Vincent, Sophie Marty, Olivier Tandonnet, Alexis Redor, Xavier Valette, Ourida Aoudia, Jacques-Arnaud Seyrig, Bertrand Beaune, Hugues Aumaitre, Georges Pinon, Yann Leveneur, Sylvie Charachon, Raoul Herbrecht, Henriette de Valk, Gary David, Julien Pouyanne, Marc Dommergues, Majed Al Chaar, Véronique Blanc-Amrane, Pascale Guillarmé Grossmann, Bruno Abraham, Yves Morel, Philippe Suel, Denis Sautereau, Olivier Guilloy, Tu Anh Tran, Frédéric Laurent, Zahir Amoura, Jacques-Olivier Bay, Zoubida Elharie-Heraux, Joyce Sibony-Prat, Bernard Guillois, Dominique Rohmer-Heitz, Audrey Barrelet, Jérémie Courouble, Jean-Paul Herry, Daniel Vittecoq, Annie Vermesch-Langlin, Jean Auroux, Claude Aubert, Thierry Harvey, Ghislaine Lamoine-Gimet, J. Riahi, Florence Soraudeau, Bachar Al-Jalaby, Caroline Périsson, Khélifa Ayouz, Florence Cardot, François Maillet, Alain Goux, Théophile Magna, Bertille de Barbeyrac, Adrien May, Dominique Andreotti, Olivier Jonquet, Hélène Dumouchel, Didier Thibaud, Philippe Morlat, Pascal Chevalet, Pascal Ancelin, Guy Chambreuil, Cécile Le Boterff, Anne Ceriez, Olivier Detante, B Pangon, Claude C.A. Bernard, Vincent Cailleaux Pierre-Etienne Cailleux, Jordi Miatello, Pierre-Yves le Berruyer, Sylvain Kouaho, Michel Briaud, Hélène Delaby, Patrick Herbecq, Christine Segonds, Véronique Jault, Pascale Brunel, Christine Dussopt, Jean Thore, Jean-Marc Thouret, Jean-Marc Kerleau, François Le Baron, Slavius Matica, Sophie Leautez-Nainville, Matthieu Pecquet, Laurent Bret, Yacine Sedjelmaci, Pierre Metton, Habiboulaye Diallo, Jany Rey Zermati, Arnaud Delahaye, Hélène Chaussade, Laurent Mandelbrot, Emilie Bessede, Olivier Casanovas, Paul Pierrot, Annick Legras, Dominique Lauque, Hélène Gatti, Jean Catineau, Ebutu Likose, Gilles Capellier, Eric Kibbrecht, Freddy Thibaut, Patrick Valadier, Chantal Lemble, Joël Gaudelus, Joelle Mellier, Joëlle Brochen, Emmanuel Gascou, Stéphane Bonacorsi, Stéphanie Bannier, Bruno Fantin, Didier Raffenot, Valérie Revel, Hakim Amroun, Huguette Negrery, Anne-Laure Fauchais, Paul Mercury, Michel Chuzeville, Christian Zumbo, Nicolas Després, Pascal Roblot, Jérôme Pasche, Jean Claude Boufetteau, Jocelyne Caillon, Julien Boileau, V. Rabier, Benjamin Manéglier, Emilie Jourdes, Franck Ceppa, Christine Recule, Nicolas Degand, Benoît Henry, Thierry Baranger, Dominique Pateron, Agnès Pélaquier, Gérard Bouchet, Hélène Fiette, Ozel Guiden, Dana Ranta, Etienne Ruppé, Nabil Chiouk, Jacques Breuil, Dominique Leduc, Véronique Loustaud, Hervé Métenier, Michel Durand, Isabelle Mahé, Leila Karaoui, Marie-José Collus, Mehran Monchi, Olivier Belmonte, Romain Blondet, Jacques Thierry, Karine Humbert, Gilles Salama, Marie-Noelle Heurtaux, Cécile Goujard, Bruno Sivery, Martial Boisseau, Redouane Dahoumane, Pierre Delour, Christian Niels Meyer, Anne Faudon-Gibelin, Gérard Poulain, Roger-Charles Luciani, J.-C. Souquet, Olivier Grossi, François Vandenesch, Sylvain Mermont, Jacques Bronner, Sonia Dahan, Paul Marzouk, Pascal Pouedras, Noureddine Djafari, François-Xavier Caroli-Bosc, Jean-François Dessin, Brigitte Gruffat, Armelle Morin-Fatome, Sylvie Thoinet, Bano Konate, Jean-Winoc Decousser, Claire Poyart, Patrick Plessis, Olivier Millet, Vincent Cattoir, Françoise Geffroy, Manica Vasseur, Pierre Carli, Isabelle Citony, Christian Richard, Nicolas Sigur, Patrick Marthelet, Luwawu Mbimba, Pierre Feugier, Philippe Sauder, Hama Djerad, Evelyne Bourgerette, Hanen Chahtour, Adrien Lemaignen, Dominique Bechade, Patrick Ochocki, Antoine Vieillard Baron, Dominique Astruc, Marie-Pierre Moiton, Nicolas Dubois, Sylvie Ledru, Corinne Seknazi, Hélène Poupet, Jean-Philippe Brieux, Gérard Barthélémy, Aihem Yehia, Louis-Jean Couderc, Ahmed, Françoise Rigaux, Yohann N'guyen, Philippe Bethery, Damien Corberand, Etienne Auvray, Paul-Louis Woerther, Christian Combe, Sophie Delesalle, Jean-Marie Piala, Faraj Al Freijat, Philippe Juvin, Malcolm Lemyze, Hyacine Rey, Claire Larible, Noel Milpied, Lémia Zgarni, Julia Gaillard, Agnès Juven, Paola Otean, Adrien Melis, André Pechinot, Olivier Bouchaud, Olivier Chassin, Pierre Hausfater, Asma Trabelsi-Jnifen, Vincent Grobost, Didier Lemery, Pierre Soury, Françoise Brevet, Jacques Tankovic, Dominique Sansot, Jean Louis Salomon, Charlotte Cordonnier, Brigitte Lamy, Antoine Maisonneuve, Dominique Pressac, Claude Rémy, Rodolphe Sobesky, Stéphanie Cognet, Pierre Cougoul, Didier Jan, Dominique Perrotin, Cécile Hombrouk-Alet, Thierry André, Gilbert Pochmalicki, Serge Girard, Vincent Zerr, Guillaume Cadiot, Claudine Lasbasses, Michel Slama, Abderrazak El Yamani, Sophie Brovedani, Jean Armengaud, Romain Hernu, Géraldine Mascade, Aurélien Lorléa'ch, Ali Akkari, Mathieu Tourdjman, Christopher Payan, Eric Jullian, Nathalie Fonsale, Frédéric Riehl, Paul Strock, Geneviève Grise, Philippe Mottaz, Christian Floriot, Marie-Noëlle Ungeheuer, Denis Caillot, Arnaud Chalvon-Demersay, Catherine Branger, Stanislas Bruley des Varannes, Marc Paccalin, Marie-Pierre Danjean, Alexandre Mebazaa, Xavier Brunet, Roland De Varax, Laurence Delhoustal, Sophie Haro, Bruno Chabanon-Pouget, Isabelle Goidin, Dominique Chudersky, Corinne Costes, Delphine Chatellier, Maud Gelez, Damien Dassant, Pascal Joly, Jean-Michel Arnal, Zakaria Hamitou, Philippe Rondepierre, Carole Pignon, Valérie Crombe, Amanda Lopes, Chrystelle Kemenar, Olivia Raulin, Anne-Cécile Hochart, Sandrine Gérart Pons, Valérie Zeller, Guillermo Reyes Ortega, Mathilde Guérin, Audrey Migraine Bouvagnet, Florence Eboue, Isabelle Loury-Lariviere, Sophie Leotard, Suzanne Lima, Marie Kassis, Jean-Luc Donay, Jean-Pierre Audié, Guillaume Cartron, Arnaud Ribier, Fanny Buron, Mirela Tuca, Marius Semenescu, Arnaud Serre, Vincent Quentin, Denise Bouyssou-Destriau, Violaine Bresson, Christine Chandesris Joséphine Chapalain-Cagnon, Eric Cua, Henri Courtade, François Bénézit, Sébastien Lamache, Philippe Bonnefoy, Francis Schneider, Richard Monarchi, Adeline Schendel, Paramasiven Mootien, Ghislaine Gardes, Pierre-François Westeel, Jean-François Magny, François Caron, Jocelyn Michon, Didier Eyer, Isabelle Ronda, Pierre-Yves Robillard, Frédéric Renou, Anna Faucher, Jean-Robert Harlé, Anne Debernardi, Grégory Akerman, Benoît Fontenel, Pierre Hourdebaigt-Larrusse, Marie-Noëlle Adam, Aude Lessene, Abdelkader Hrichi, François Blot, Athéna Le Pierres, Romain Lemarie, Françoise Granier, Véronique Tardy, Marc Gatfosse, Pierre-Marie Roger, François Goupil, Saïd Aberrane, Franck Bernardi, Isabelle Plantier, Nathalie Funakoshi, Jean-Gilles Delecalle, Patricia Barbut, Jacques Reynes, Christophe Roy, Sophie Perreve, Michel Garre, David Ribes, Cyrille Ede, Jean-Claude Dausset, Francis Duchene, Jean Caussin, Michelle Becker-Schneider, Gilles Berthelot, Damien Dupont, Jean-Michel Gillot, Aurélie Messager, Jean-Marie Pannecouck, Jean-Christian Roussel, Alain Reynaud, Sylvie Cariou, Anne Dao, François Guillemot, Martin Martinot, Patrick Casali, Anne-Sophie Poirier, Aissa Kerchache, Necera Sakek, Eric Porthault, Christophe Decoene, Chantal Ache-Papillon, Brigitte Bicais, Jean-Claude Feugier, Thierry Masseron, Charles Marty-Ane, Daniele Goldgran Toledano, Jean-Christophe Dubus, Damien du Cheyron, Dominique Decré, Jean-Loup Pennaforte, Ahmed Tigaizin, Bernard Vache, Eric Oswald, Claire Moulinoux, Anne-Christine Jaouen, Caroline Charlier, Anne-Laure Virlouvet, Ali Kara, Jean-Luc Sicsic, Sylvie Goffart, Mathieu Zuber, Claudine Fèbre, Olivier Lortholary, Mathieu Dupont, Annie Vessieres, Thierry Helvadjian, Thomas Signouret, Cedric Daupin, Sandrine Essouri, Jean-Louis Jacob, Pascal Boileau, Caroline Blazejewski, Quentin Lepiller, Juan-Pablo Maureira, Eddy Lebas, Christophe Deschamps, Amévi Ananivi, Clovis Foguem, Daniel Adoue, Abdourahim Chamouine, Alain Michault, Bruno Guérin, Olivier Baud, Clara Vinci, Thierry Weitten, Jean-Marc Eychene, Marie Froidure, Julien Obiols, Patricia Roussellier, Marc Lecuit, André Cabié, Saskia Foucart, Karim Belhadj, Michel Cingotti, Bruno Dumoulard, Jean Puyhardy, Etienne Danquechin Dorval, Lucile Mendes-Martin, Enrique Casalino, Luc Jarrige, Fabien Lambiotte, Philippe Masson, Mohammed Mansouria, Pierre Thouvenot, Katy Jeannot, Martine Nyunga, Valérie Macci, Florent Masia, Claire Briere de la Hosseraye, Wassila Anteur, André Sommabère, Marie-Claude Germain, Isabelle Arnault, Bernard Carbonelle, Philippe Devos, Daniel Protar, Tiphaine Gaillart, Ludovic Lassel, Laurence Hamou-Plotkine, David Trystram, Thierry Bureau, Olivier Collard, Fanny Vuotto, Sophie Malhiere, Frederique Canis, Gillles Plainfosse, Catherine Lechiche, Bertrand Lassere, Martine Chouraqui, Jean Baptiste Michot, Fethi Radhouane-Khanjari, Carole Barbier, Pascal Bonitchi, Abdelaziz Benkhelil, Odile Salmon, Laurent Damaj Gandhi, Bertrand Minguet, Michel Wagner, Odile Falguières, Zahr-Eddine Ali Chaouche, Eric Zaoui, Isabelle Guichard, Bernard Huttin, Apollinaire Karirisi, Gaël Cinquetti, Christophe Plane, Lionel Rostaing, Yanne Henry-Andrieu, Daniel Re, Virginie Verrier, Pascal Bolot, Michel De Biasi, Laurence Vrigneaud, Mathilde Turpin, Marie-Claude Demachy, Etienne Roussel, Michèle Blancs, Olivier Join-Lambert, Yves Ville, Thierry Granger, Gilles Hilbert, Virginie Medeau, Daniel Villers, Benoit Pilmis, François Gouraud, Emmanuel Ardiet, Catherine Heyraud-Blanchet, Alain Devidas, Hélène Dieye, Julie Cremniter, Jean-François Bergmann, Rozenn Le Berre, Virginie Leguen, Daniel Royer, Gilles Le Maout, Christian Harou, Sylvie Gabriel-Soléan, Yves Regouby, Martine Pestel-Caron, Patrick Brunet, David Boutoille, Emmanuelle Bonnin, Patrice Coulon, Marc Sullice, Marianne Barbieux, Gilles Cambonie, Joëlle Tricoire, Marie-Nadège Bachelier, Delphine Briend, Céline Ramanantsoa, Nathalie Bednarek, Didier Lebreton, Julien Lagrandeur, Damien M'Bey, Philippe Audeguy, Elie Saliba, Lena Damaj, Hassan Fallouh, Pascal Couturier, Fabrice Prévost, Yves Domart, Marie-Odile Lafforgue, Anne Le Du, C. Beuscart, Pierre Guillet, Fabrice Larrazet, Marie-Hélène Hausermann, Henri Robert, Nicolas Fanjaud, François Goehringer, Thomas Bachelot Philippe Badia, Jean-Michel Coulaud, Cristel Fissore Magdelein, Renaud Defebvre, Anne-Sohie Moreau, Johan Courjon, Gilles Salles, Michel Mialon, Silvia Iacobelli, Emmanuelle Bille, Marie-Christine Barbier, Yves Aubard, Patrice Badila, Jean-Philippe Rasigade, Alban Deroux, Evelyne Lecaillon-Thibon, Michel Godin, Abdelmajid Djeffal, Viorica Badurescu, Meriem Canitrot, Pierre Blanchard, Antoine Legros, Laurence Got, Françoise Duluc, Mylène M. Maury, Gilles Dassieu, Nordine Khodeir, Jean-Marie Duez, Mathieu Morincomme, Jérôme Lacroix, Mathieu Revest, Koffi Blewoussi, Isabelle Barazer, Françoise Poitevin, Camille Seignovert, Stéphanie Honore Bouakline, Anne Heidt, Brigitte Malbruny, Julien Desblache, Christian Cattoen, Eric Jaunait, Bruno Chaminade, Claude Bazin, Jonathan Chelly, Anne Pottier, Alain Schmitt, Alain Tissot, Karim Dadoun, P. Rebattu, Claudine Contamin, Arnaud Guerard, Nathalie Ravet, Sandrine Khalifa-Thellier, Marlène Chatron, Gaëlle Dörr, Hélène Biessy, Emmanuel Forestier, Bruno Devaux, Jean-Jacques Grelaud, Xavier Tchenio, Marie-Cécile Ploy, Jérémie Violette, Michèle Burdin, Lionel Falchero, Dominique Jacomy, Jean-Christophe Rozé, Damien Labarriere, Stéphane Leroux, Corinne Meregnani, Assia Ferhat Carre, Paul Orode, Jean-Gabriel Paul, Catherine Godon, Agnès Vinay, Régine Barraduc, Dominique Dallay, Alexandre Ampère, Anne-Gaelle Kervegant, Guillaume Louart, Dominique Beal Ardisson, Francoise Leonetti, Jean-Yves Baril, Stéphanie Haiat, Bincy Darre, Jérôme Bay, Yvan Gauthier, Sylvie Radenne, Pierre-Yves Gueugniaud, Philippe Ravaud, Luc Landraud, Guillaume Ranchon, Loïc Chimot, Véronique Duval, Ilhem Agha-Mir, Sabine Camiade, Estelle Wafo, Jean-Patrick Laporte, Mariam Roncato-Saberane, Camille Bron, Patrice Laudat, Samir Kennouche, Nawel Afroukh, Dominique Neri, Hakim Kherouf, Yoar Hichri, Pierre-Edouard Bollaert, Gwenaelle Vary, Denis Castaing, Christine Lefort, Sébastien Rouget, René Robert, Christelle Guillet-Caruba, Catherine Simonin, Alain Vighetto, Severine Cabasson, Alain Brusset, Alexandra Doloy, Christel Cherlet, Ahmed Rouidi, Marina Salvucci, Réginald Pordes, René-Gilles Patrigeon, Emmanuelle Dupre-Narlet, Jacques Minet, Fethi Taleb, Anne-Marie Colingorski, Tahar Hadou, Sylvain Diamantis, Isabelle Glorieux, Thierry May, Jean-Claude Colombani, Anne Berth-Farges, Nicole Desplaces, Renaud de Tayrac, Elisabeth Walter, Fabienne Lorge, Pascal Reboul, Nathalie Dournon, Laurence Estépa, Marie-Lina Toubia, Mathilde Flahault, Thierry Delacour, Dominique Hurel, Hélinoro Andriamaneo, Cécile Bébéar, Denis Grasset, Miloud Serier, Oléna Orléva, Nadine Dubroca, Hervé Gentilhomme, Jean-Luc Baudel, Isabelle Lavenu, Salim Smati, Carlo Saroufim, Eric Placidi, Albert Sotto, Benoît Libeau, Hélène Leroy, François Golfier, Christophe Dollon, Laurence Desnoulez, Eric Barre, Daniel Cohen, Pascal Priollet, Thierry Marsepoil, Benoît Lionnet, Jacques Tebib, Pascale Penn, Antoine Bouissou, Christian Roth, Olivier Martinet, Anna Schmitt, Nathalie Fruleux, Fouzia Radaoui, Jean-Marc Lessinger, Virginie Morando, Jean-Jacques Maillet, Christophe Fruchart, François Boué, François Goffinet, Franck Lellouche, Martin Demarchi, Alain Geissler, Jean-Charles Picaud, David Assouline, Patricia Brazille, Philippe Guimier, Marie-Françoise Dabysing, Bruno Delpeuch, Vanessa Tran, Guy Gengembre, Delphine Deligne, Dominique Vodovar, Yvan Touze, Sabrina Parent, Anne Decoster, Camille Dewitte, Emmanuel Weiss, Thierry Lambert, Thomas Guimard, Vincent Caille, Claude Guérin, Françoise Evreux, Geneviève Barjon, Basile Ondze, Damien Fournier, Olivia Bandin, Sophie Mignart, Henri Demontclos, Didier Perez, Jacques Croize, Nicole Desbois-Nogard, Guenièvre Imbert, Clarisse Dupin, Khalid Ridah, Marie-Christine Varin, Guillaume Arlet, Edith De Clareuil, Marie-Line Eustache, Patricia Le Pimpec, Louise Fortin, Eugène Ngami, Fabrice Mihout, Cecilia Esnault, Vincent Bouden, Véronique Annaix, Yves Poinsignon, Aurélien Lorchleac'h, Jean-Marc Degreff, Marie Garofano, Renaud Mesnage, Anne-Marie Roque-Afonso, Alain Chevailler, Stéphane Hominal, Thierry Charbonnier, Adrianna Bildea, Fabien Fily, Benjamin Davido, Emmanuel Rassiat, Assi Assi, Stéphanie Brunet, Hervé Jacquier, Catherine Claise, Annie Durand, Yannick Monceau, Pierre Blanc, Jean-Marie Sire, Yves Sucin, Jean-Pierre Zarski, Nathalie Bronet, Ingrid Lafon, Philippe Rey, Jacques Markarian, Eric Sennevile, Olivier Wink, Guilène Barnaud, Anne-Sophie Peultier, Sabine Taylor, Rim Savatier, Patrick Valayer, Claude Negrier, Selim Jennane, Edouard Begon, Laura Hyerle, Delphine Bridoux, Claire Daurel, Benoît Dalle, Mathilde Lescat, Philippe Stolidi, Elodie Perrodeau, Xavier Heches, Pierre Castelnau, Philippe Bray, Jean-Claude Texier, Serge Rossignol, Maud Brung-Lefebvre, Jean-François Subra, Jean-Marie Delarbre, Morgane Schneerson, Guyro Jang, Mona Mehri, Nathalie Landgraf, Pierre-Marie Girard, Armand Goll, Zaineb Bekguesmia, Christophe Clement, Michel Collet, Vincent Maze, Amine Benjelloul, Solène Durliat-Ellie, Vincent Letouzey, François Schmitt, Valérie Martinez, Sarah Watson, Abderrezak Bouasria, François Barbier, Raphael Lauretta, Mirana Razafimahery, Cristina Sirbu, Patrick Malherbe, Anne Wuillai, Ludovic Lesecq, Philippe Gaudard, Serge Houssaye, Jacques Monsegu, Gilles Rival, Chantal Chaplain, Jean-Didier Grangé, Oana Zamfiri, Florence Nguyen-Khac, Marc Portneuf, Jean-Michel Pawlotsky, Delphine Bonnet, Laurent Traissac, Sophie Hamon-Charles, Didier Dreyfuss, Louis Bernard, Laurence Detourmignies, Olivier Martineau, François Pettinelli, Marc Zandecki, Michel Dreyfus, Alain Chapelle, Sébastien Sabbat, Anne-Sophie Labussiere, Jean-Louis Gaillard, Chloé Plouzeau-Jayle, Patrick Zoveda, Véronique Leflon, Marie Levy, Aurélie Labé, Bruno Soulie, Raoul Jacques Bensaude, Hecham Moussa, Sylviane Catteu, Nathalie Biron, Loïc Masson, Georges Mourad, Nejla Aissa, Dragos Ciocan, Hubert De Boysson, Jean-Luc Bouyer, Patrick Yeni, Thierry-Pascal Zame, Caroline Thomas, François Cavalié, Laurence Koulmann, Christophe Rioux, Olivier Barraud, François Bricaire, Marguerite Le Poulain, Marie-Noelle Noulard, René Thomas, Guy Semet, Laurent Mosser, Olivier Marret, Brigitte Rivière, Vincent Jarlier, Jean-Philippe Coindre, Marc Villemain, Martin Pierre, Yacine Benkaci, Philippe Chiron, Hoang Vu-Thien, Jérôme Gournay, Andrea Labaune-Kiss, Brigitte Lauzanne, Fanny Lemercier, Souad Silhadi, Imad Kansau, Christophe Poncelet, Olivier Baldesi, Francis Thuet, Olivier Leroy, Aurore Lamberet, Camille Petit Hoang, Sophie Micheli, Ayman Abokasem, Hakima Nesrine, Pierre Lureau, Christian Chidiac, Vincent Piriou, Fabien Zoulim, Dieudonné Nicobaharaye, Anne Tixier, Isabelle Matheron, Soumeth Abasse, Victoria Cacheux, Serge Herson, Christine Fuhrmann, Olivier Proost, Bernard Bedock, Olivier Rogeaux, Mostapha Hajjar, Anne Reverseau, René Courcol, Françoise Carmagnol, Yves Guénard, Céline Ménard, Bouchra Lamia, Bruno Lemmens, Damien Bouhour, L. Lequen, Gaëlle Baty, Cédric Bouet, Dominique Guerrot, Stéphane Blanc, Catherine Chirouze, Anne-Hélène Reboux, A. Vachée, Gregory Taurin, Myriam Mein-Bottini, Jean-Pierre Belot, Alain Lafeuillade, Patricia Gabez-Therou, Philippe Labrousse, Bernard Jarrousse, Philippe Noto, Vincent Brunot, Philippe Condominas, Marion Challier, Béatrice Berçot, Delphine Anuset, Mélanie Daval Cote, François Bernasconi, Y. Costa, Chandrah Goburdhun, Bernard Gressier, Alban Michaud-herbst, Franck Charlier, Moussa Hecham, Luc Boulain, Hélène Corneloup, Alix Greder Belan, Nicolas Boussekey, Claire-Antoinette Dupuy, Yannick Rouquet, Benoit Renard, Benifla Jl, Etienne Javouhey, Michèle Granier, Marie-Christine Jaffarbandjee, Emilie Piet, Benoît Bergues, Claire Malbrunot, Catherine Tiry, Philippe Mérigot, Mouna Ben Soltana, Chantal Roure Sobas, Florian Radenac, Yves Thomas, Agathe Blaise, Sylvie De Martino, Laurence Legout, Gabriel Choukroun, Jean-François Muir, Peggy Dupretz, Patrick Dupont, François Guichart, Julie Jean, Jean-Michel Descamps, Bernard Kittschke, Anne Gruson, Gerard Viquesnel, Marie Keller, Pascal Chavanet, Françis Vallet, Yvan Vaschalde, Jean-Luc Hanouz, Gerard Lina, Françoise Loison, Simon Vincent, Jean-Paul Thellier, Moncef Afi, Dominique Zagozda, Hélène Sokeng-Affoule, Marc Le Bideau, Jean-François Loriferne, Alain Gravet, Sophie Deprecq, Tarik Naceur, Severine Mielczarek, René-Jean Bensadoun, Bernard Karkous, Yves Bléher, Jocelyne Poulain, Véronique Goulet, Laurence Nicolet, Sophie Arista, Antônio Lúcio Teixeira, Jean-François Schved, Laurent Nicolet, Claire Lecomte, Faiza Benddif-Fin, Michel Aumersier, Laurence Burc-Struxiano, Maxime Thouvenin, Samia Harbi, Mathieu Detave, Catherine Rebeyrotte, Jean-Paul Kisterman, Bruno Berdin, Pascal Vincent, Laurent Argaud, Elisabeth Parisi-Duchene, Geneviève Julienne, Fernanda Farto-Bensasson, Georges-Fabrice Blum, Sad Gaizi, Tali-Anne Szwebel, Raphaël Lepeule, Marie-Thérèse Climas, Anne-Françoise Dillies, Amar Boudhane, Umberto Simeoni, Pierre-François Dequin, Gérard Oliviero, Alain Gourlaouen, Caroline Piau, Marie-France Lutz-Murphy, Benoît Claude, Jean-Paul Aubry, Nadine Dubosc-Marchenay, Kamilla Chraibi, Emmanuelle Heusse, Sylvain Le Chevallier, Nathalie Brieu, Farid Sifaoui, Lorraine Letranchant, Hélène Durox, Jean-Pierre Lagasse, Adel Ghedira, Xavier Roubert, Fatma Magdoud, Hélène Jean-Pierre, Etienne Carbonelle, Olivier Dereeper, Lionel Carbillon, Christophe Billy, Mélanie Roblin, Marie-José Kodzin, Philippe Niel, Solène Makdessi, Matteo Vassallo, Maryse Archambaud, Fabian Haccourt, Didier Blaise, Stéphane Bourgeois, Elena Marcu, Charles Kubiak, Brisse Castel, François Guinet, Marie Pouzoullic, Frédérique Nathan-Bonnet, Vincent Gendrin, Céline Becherrawy, Aline Secher, Pierre Abgueguen, Clarence Eloy, Jean-Marc Tourani, Frédéric Klapczynski, Bernard Montmasson, Philippe Real, Joanna Pofelski, Yves Welker, Karim Krechiem, Eric Caumes, Martine Elena-Daumas, Christophe Saigne, Gilles Hittinger, Chantal Delesalle, Jonathan Messika, Fabrice Lesage, Daniela Pop, Daniel Coetmeur, Renato Colamarino, Chetaou Mahaza, Patrick Plésiat, Isabelle Fredenucci, Mylène Baret, Guy Mager, Pascale Chavel, Isabelle Labourdette, Anne-Claude Menguy, Nicolas Fortineau, Ludovic Le Sec, Valérie Gauduchon, Francis Barraud, Nicolas Letellier, Didier Vincent, Janine Frey, Philippe Riegel, Michel Pavic, Jean-Luc Fabre, Jean-Pierre Fauchart, Alain Goudeau, Stéphanie Husson-Wetzel, Philippe Eymerit, Mohamed Camara, Nathalie Seta, Elisabeth Carole Ngo Bell, Philippe Repellin, Laurent Alric, Vincent Leroy, Françoise Delisle Mizon, Jean-Philippe Emond, Marie-Françoise Borie, Lise Crémet, Wladimir Chelle, Elisabeth Brottier-Mancini, Bernard Garrigues, Claire Letellier, Loïc Geffray, Frédéric Méchaï, Julien Bador, Benoit Guery, Alain-Charles Fouilhoux, Corinne Dagada, Pierre Duhaut, Julien Goustille, Arnaud Sément, Francis Carcenac, Isabelle Girard-Buttaz, Claire Chapuzet, Fabienne Jouatte, Bruno Riou, Fabrice Hayoun, Chloé Di Meglio, Youssef Ali, Michel Leneveu, Nathalie Montagne, Yves Garcera, Audrey Moustache, Pierre-Eric Danin, Geneviève Le Lay, Dominique Courouge-Dorcier, Isabelle Worcel, Emmanuel Morelon, Vincent Pestre, Jean-Pierre Vilque, Jean-Christophe Paquet, Lucien Bodson, Anne-Marie Forest, Fabrice Pierre, Christian Pommier, Fabien Dutasta, Pierre Fournel, Stéphanie Courtois, Elodie Dubois, Serge Vanden Einjden, Patrick Honderlick, Pascal Richette, Fabienne Tamion, Véronique Chassy, Richard Megbemado, Anne-Marie Le Reste, Bernard Simian, Henri Osman, Anthony Texier, Badih Ayach, François Simon, Jean-Michel Filloux, Béatrice Dubourdieu, Jean-Claude Semet, Sarah Kubab, Tawfiq Henni, Patrick Dudeffant, Delphine Hequet, Olivier Mimoz, Marc Auburtin, Amélie Chabrol, Mickael Bonnan, Caroline Léonnet, Claire Wintenberger, Serge Ilunga, Patrice Lanba, Sophie Rosello, Alexandre Damage, Flore Bouche, Michel Thibault, Frederic Faibis, Chantal Dhennain, Jean-Philippe Talarmin, Armelle Lamour, Remi Boussier, Fabien Garnier, Marie-Laure Brival, Nourredine Hedjem, Philippe Vande-perre, Raphaël Coint, Jean-Claude Reveil, Eva Weinbronn, Emmanuelle Lavalard, Alexandra Fille, Françoise Le Turdu, Lionel Leroux, Jean-Yves Lefrant, Jean Berthet, Radia Bouaziz, Alain Ravaud, Sylvaine Rousseau, Yacine Merrouche, Alain Le Coustumier, Bertrand Guider, Gisèle Dewulf, Jean-Marc Faucheux, Jacques Piquet, Franck Leibinger, Charles Cerf, Robin Stephan, Jean-Philippe Redonnet, Jean-Paul Stahl, Ella Dzeing, Simona Pavel, Guy Vernet, Ghada Hatem, Samer Kayal, Jacques Deschamps, Dominique Descamps, Marion Levast, Marc Bouiller, Sylvie Dargere, Claire Dingremont, Stéphane Gaudry, François Maillot, Sylvie Odent, Nathalie Cervantes, Hélène Zanaldi, Laurence Gachassin, Olivier Ruyer, David Patin, Benoît Cazenave, Pascal Jacquier, Michelle Boyer, Béatrice Berteaux, Virginie Zarrouk, Jacques Bor, Isabelle Legoff, Hélène Albinet, Florence Rousseau, Gilles Pialoux, Guenaelle Salaun-Beretta, Alexandra Moura, Véronique Vernet Garnier, Didier Lepelletier, Pierre-Alexandre Hauss, Joëlle Belaisch-Amart, Didier Lepeletier, Jacob Xavier, Aline Nare, Annie Motard-Picheloup, Alain Améri, Bertrand Lioger, Jean-Valère Malfuson, Centre d'infectiologie Necker-Pasteur [CHU Necker], Institut Pasteur [Paris] (IP)-CHU Necker - Enfants Malades [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Centre National de Référence Listeria - National Reference Center Listeria (CNRL), Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre collaborateur de l'OMS Listeria / WHO Collaborating Centre Listeria (CC-OMS / WHO-CC), Institut Pasteur [Paris] (IP)-Organisation Mondiale de la Santé / World Health Organization Office (OMS / WHO), CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Université Pierre et Marie Curie - Paris 6 - UFR de Médecine Pierre et Marie Curie (UPMC), Université Pierre et Marie Curie - Paris 6 (UPMC), Département de Médecine interne [Lariboisière], Hôpital Lariboisière-Fernand-Widal [APHP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Diderot - Paris 7 (UPD7), Biologie des Infections - Biology of Infection, Service de Gynécologie et Obstétrique [Cochin], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Investigation Clinique et d’Accès aux Ressources Biologiques (Plate-forme) - Clinical Investigation and Access to BioResources (ICAReB), Institut Pasteur [Paris] (IP), Infectious Disease Department [Saint Maurice], Agence Nationale de la Santé Publique [Saint-Maurice] (ANSP), Centre de Recherche Épidémiologie et Statistique Sorbonne Paris Cité (CRESS (U1153 / UMR_A_1125 / UMR_S_1153)), Institut National de la Recherche Agronomique (INRA)-Université Paris Diderot - Paris 7 (UPD7)-Université Paris Descartes - Paris 5 (UPD5)-Université Sorbonne Paris Cité (USPC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris Descartes - Paris 5 (UPD5), CHU Amiens-Picardie, Mécanismes physiopathologiques et conséquences des calcifications vasculaires - UR UPJV 7517 (MP3CV), Université de Picardie Jules Verne (UPJV)-CHU Amiens-Picardie, MONALISA study group, Programme Hospitalier Recherche Clinique, Institut Pasteur, Inserm, French Public Health Agency., ROZIER, marie-Claire, CHU Necker - Enfants Malades [AP-HP], Centre National de Référence Listeria - Biologie des Infections (CNRL), Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre collaborateur de l'OMS Listeria - Biologie des Infections (CCOMS), CHU Pitié-Salpêtrière [APHP], Hôpital Lariboisière-Université Paris Diderot - Paris 7 (UPD7), Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-CHU Cochin [AP-HP], Institut Pasteur [Paris], Assistance publique - Hôpitaux de Paris (AP-HP) (APHP), Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Necker - Enfants Malades [AP-HP], Centre National de Référence Listeria - Biologie des Infections ( CNRL ), Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Centre collaborateur de l'OMS (CCOMS) des Listeria ( CCOMS ), Centre de Recherche Épidémiologie et Statistique Sorbonne Paris Cité ( CRESS (U1153 / UMR_A 1125) ), Institut National de la Recherche Agronomique ( INRA ) -Université Sorbonne Paris Cité ( USPC ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Université Pierre et Marie Curie - Paris 6 - UFR de Médecine Pierre et Marie Curie ( UPMC ), Université Pierre et Marie Curie - Paris 6 ( UPMC ), Université Paris Diderot - Paris 7 ( UPD7 ) -Hôpital Lariboisière, Biologie des Infections, Institut Pasteur [Paris]-Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Investigation Clinique et d’Accès aux Ressources Biologiques (Plate-forme) - Clinical Investigation and Access to BioResources ( ICAReB ), Agence Nationale de la Santé Publique [Saint-Maurice] ( ANSP ), Assistance Publique - Hôpitaux de Paris, Assistance publique - Hôpitaux de Paris (AP-HP), Université Paris Descartes - Paris 5 ( UPD5 ), Institut Pasteur [Paris]-CHU Necker - Enfants Malades [AP-HP], Institut Pasteur [Paris]-Organisation Mondiale de la Santé / World Health Organization Office (OMS / WHO), Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), and Université Paris Diderot - Paris 7 (UPD7)-Université Sorbonne Paris Cité (USPC)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de la Recherche Agronomique (INRA)
Subjects: Bacteremia/epidemiology/mortality, 0301 basic medicine, Male, Pediatrics, bacteraemia, Infectious Disease Transmission, [SDV]Life Sciences [q-bio], Bacteremia, France/epidemiology, Infant, Newborn, Diseases, Foodborne Diseases, Meningoencephalitis, Pregnancy, Risk Factors, Vertical, Medicine, Listeriosis, Prospective Studies, Pregnancy Complications, Infectious, Prospective cohort study, ddc:618, diabetes, alcoholism, Hazard ratio, Foodborne Diseases/microbiology, immuno suppressive therapies, Prognosis, 3. Good health, [SDV] Life Sciences [q-bio], Hospitalization, Infectious Diseases, isolates, Population Surveillance, Female, France, Listeria monocytogenes/classification/isolation & purification, Cohort study, Adult, medicine.medical_specialty, 030106 microbiology, Notifiable disease, Listeriosis/diagnosis/epidemiology/microbiology, Context (language use), macromolecular substances, 03 medical and health sciences, Humans, study, Aged, [ SDV ] Life Sciences [q-bio], business.industry, Public health, cirrhosis, Infant, Newborn, Infant, Diseases/epidemiology/microbiology, HIV, Mandatory Reporting, Newborn, medicine.disease, Listeria monocytogenes, infection, Infectious Disease Transmission, Vertical, Pregnancy Complications, Infectious/epidemiology/microbiology, Meningoencephalitis/epidemiology/microbiology/mortality, Observational study, business, prognostic, mellitus
Abstract: International audience; Evidence before this study We searched PubMed on June 30, 2016, for English-language cohort studies published since Jan 1, 1980, of patients with invasive listeriosis worldwide with the keywords " listeria " , " listeriosis " , " maternal " , and " neurolisteriosis ". Studies had to include epidemiological or clinical data on listeriosis. All clinical forms of infection were included (bacteraemia, neurolisteriosis, and maternal–neonatal infection). Host risk factors for listeriosis have been well identified, but the clinical features and prognostic factors of the disease are based on retrospective studies compiling heterogeneous data or random collected cases. Furthermore, no clinical trial has ever been done and medical management is not evidence based. Added value of the study Our study is the first prospective clinical study focusing on all forms of invasive listeriosis. The study is based on a national mandatory system that allowed the nearly complete capture of microbiologically proven cases. The study shows a higher burden of listeriosis than reported before: more than 80% of infected mothers experienced major fetal or neonatal complications (fetal loss, very high prematurity, early or late onset disease); only 39% of patients with neurolisteriosis survived and fully recovered. The study provides important new data to improve management and predict outcome in listeriosis, such as determination of the time window for fetal losses (
Published: 2016
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38. Expression of complement and toll-like receptor pathway genes is associated with malaria severity in Mali: a pilot case control study

Author: Guy Vernet, Amadou Niangaly, Abdoulaye Djimde, Rafal S. Sobota, Christopher V. Plowe, Philippe Leissner, Ogobara K. Doumbo, Mahamadou A. Thera, Jason A. Bailey, Abdoulaye K. Kone, Jessica E. Manning, Scott M. Williams, and Antoine Dara
Subjects: Male, 0301 basic medicine, Plasmodium, Microarray, media_common.quotation_subject, Plasmodium falciparum, 030231 tropical medicine, Pilot Projects, macromolecular substances, Biology, Mali, Parasite load, GZMB, 03 medical and health sciences, 0302 clinical medicine, parasitic diseases, medicine, Cluster Analysis, Humans, Malaria, Falciparum, Oligonucleotide Array Sequence Analysis, media_common, Molecular Epidemiology, Research, Gene Expression Profiling, Convalescence, Toll-Like Receptors, Case-control study, Infant, Complement System Proteins, medicine.disease, biology.organism_classification, Malaria, 3. Good health, 030104 developmental biology, Infectious Diseases, nervous system, Genetic Epidemiology, Case-Control Studies, Child, Preschool, Immunology, TLR4, Female, Parasitology, Biomarkers
Abstract: Background The host response to infection by Plasmodium falciparum, the parasite most often responsible for severe malaria, ranges from asymptomatic parasitaemia to death. The clinical trajectory of malaria is influenced by host genetics and parasite load, but the factors determining why some infections produce uncomplicated malaria and some proceed to severe disease remain incompletely understood. Methods To identify molecular markers of severe falciparum malaria, human gene expression patterns were compared between children aged 6 months to 5 years with severe and uncomplicated malaria who were enrolled in a case–control study in Bandiagara, Mali. Microarrays were used to obtain expression data on severe cases and uncomplicated controls at the time of acute disease presentation (five uncomplicated and five severe), 1 week after presentation (three uncomplicated and three severe) and treatment initiation, and in the subsequent dry season (late convalescence, four uncomplicated and four severe). This is a pilot study for the first use of microarray technology in Mali. Results Complement and toll-like receptor (TLR) pathways were differentially expressed, with severe cases showing higher expression of the C1q, TLR2, TLR4, TLR8, and CR1 genes. Other genes previously associated with malaria pathogenesis, GZMB, FOS and HSPA6, were also higher among severe cases. TLR2, TLR4, TLR8, CR1, GZMB, FOS, and HSPA6 genes were expressed at lower levels in severe cases at late convalescence. Conclusions Overexpression of genes previously associated with uncomplicated malaria was associated with severe disease. Low baseline expression of these genes may represent candidate markers for severe malaria. Despite the small sample size, results of this pilot study offer promising targets for follow-up analyses. Electronic supplementary material The online version of this article (doi:10.1186/s12936-016-1189-6) contains supplementary material, which is available to authorized users.
Published: 2016
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39. Potential implication of new torque teno mini viruses in parapneumonic empyema in children

Author: Etienne Javouhey, Lili Ren, Jianwei Wang, Li Yongjun, Gláucia Paranhos-Baccalà, Jean-Noel Telles, Alain Rajoharison, Nathalie Richard, Sandra Dollet, Guy Vernet, Johanna Galmès, Laboratoire des Pathogènes Emergents, Fondation Mérieux, Université de Lyon, Immunité infection vaccination (I2V), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-IFR128-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut des sciences cognitives Marc Jeannerod - Centre de neuroscience cognitive - UMR5229 (CNC), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS), Unité Mixte de Recherche Epidémiologique et de Surveillance Transport Travail Environnement (UMRESTTE UMR T9405), Université de Lyon-Université de Lyon-Institut Français des Sciences et Technologies des Transports, de l'Aménagement et des Réseaux (IFSTTAR), Department of Geological Sciences, University of Michigan [Ann Arbor], University of Michigan System-University of Michigan System, Emerging Pathogens Department, and BIOMERIEUX
Subjects: Male, Pleural effusion, [SDV]Life Sciences [q-bio], MESH: Base Sequence, 0302 clinical medicine, MESH: Child, Prevalence, Prospective Studies, MESH: Phylogeny, Child, Pathogen, Phylogeny, Immunoassay, MESH: Cytokines, 0303 health sciences, MESH: Chemokines, MESH: Infant, 3. Good health, MESH: Nucleic Acid Conformation, MESH: Torque teno virus, MESH: HEK293 Cells, Child, Preschool, Cytokines, Original Article, Female, Chemokines, MESH: Immunoassay, MESH: Pleural Effusion, Pulmonary and Respiratory Medicine, MESH: Cell Line, Tumor, MESH: Empyema, Adolescent, Molecular Sequence Data, Pneumonia, Viral, Biology, Virus, 03 medical and health sciences, Open Reading Frames, Immune system, Cell Line, Tumor, medicine, Humans, Empyema, MESH: Prevalence, 030304 developmental biology, MESH: Adolescent, Torque teno virus, MESH: Humans, MESH: Molecular Sequence Data, Torque teno mini virus, Base Sequence, MESH: Child, Preschool, Infant, MESH: Open Reading Frames, Pulmonary Infections, medicine.disease, Virology, MESH: Male, MESH: Prospective Studies, MESH: DNA, Viral, Pleural Effusion, Pneumonia, HEK293 Cells, MESH: Pneumonia, Viral, 030228 respiratory system, Viral replication, DNA, Viral, Nucleic Acid Conformation, MESH: Female
Abstract: International audience; An unexplained increase in the incidence of parapneumonic empyema (PPE) in pneumonia cases has been reported in recent years. The present study investigated the genetic and biological specifications of new isolates of torque teno mini virus (TTMV) detected in pleural effusion samples from children hospitalised for severe pneumonia with PPE. A pathogen discovery protocol was applied in undiagnosed pleural effusion samples and led to the identification of three new isolates of TTMV (TTMV-LY). Isolated TTMV-LY genomes were transfected into A549 and human embryonic kidney 293T cells and viral replication was assessed by quantitative real-time PCR and full-length genome amplification. A549 cells were further infected with released TTMV-LY virions and the induced-innate immune response was measured by multiplex immunoassays. Genetic analyses of the three TTMV-LY genomes revealed a classic genomic organisation but a weak identity (
Published: 2012
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40. Evaluation of Twelve Real-Time Reverse Transcriptase PCR Primer-Probe Sets for Detection of Pandemic Influenza A/H1N1 2009 Virus

Author: Wei Wang, Guilan Lu, Jianwei Wang, Yaowu Yang, Guy Vernet, Li Yongjun, Fang Huang, Qi Jin, and Richard Gonzalez
Subjects: Microbiology (medical), Neuraminidase, Hemagglutinin (influenza), Hemagglutinin Glycoproteins, Influenza Virus, Biology, medicine.disease_cause, Sensitivity and Specificity, H5N1 genetic structure, Virus, Viral Proteins, Influenza A Virus, H1N1 Subtype, Virology, Influenza, Human, Influenza A virus, medicine, Humans, Letters to the Editor, Gene, DNA Primers, Reverse Transcriptase Polymerase Chain Reaction, Molecular biology, Reverse transcription polymerase chain reaction, Molecular Diagnostic Techniques, biology.protein, Primer (molecular biology)
Abstract: Real-time reverse transcriptase PCR (rRT-PCR) assays have greatly contributed to the detection, control, and prevention of the pandemic influenza A/H1N1 2009 virus. To improve the rRT-PCR assays for detection of pandemic influenza A/H1N1 2009 virus, we evaluated the sensitivity, specificity, and performance of 12 rRT-PCR primer-probe sets [SW (a) to SW (l)] using a panel of virus strains and clinical specimens. These primer-probe sets were derived from published work and designed for detecting the hemagglutinin (HA) or the neuraminidase (NA) gene of the pandemic influenza A/H1N1 2009 virus. A primer-probe set, SW (CDC), developed by the Centers for Disease Control and Prevention (U.S. CDC) to target the HA gene of pandemic influenza A/H1N1 2009 virus, was used as a referee method. Our results demonstrated that although all primer-probe sets in this study had as high as 98.4 to 100% in silico coverage, some of the primer-probe sets had better specificity, sensitivity, and amplification efficiency than others. Two primer-probe sets, SW (h) and SW (l), which target the NA gene of pandemic influenza A/H1N1 2009 virus, were highly sensitive (10 4 copies/reaction), had high detection rates (56/60, P = 0.134, and 59/60, P = 1.000), and showed ideal specificity compared with SW (CDC). In addition, a cocktail of primer-probe sets targeted to the HA and NA genes displayed higher detection sensitivity than primer-probe sets targeting HA or NA alone, indicating that for practical applications, a combination of primer-probes targeting HA and NA genes is the best option for the detection of pandemic influenza A/H1N1 2009 virus.
Published: 2011
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41. Development and validation of a commercial real-time NASBA assay for the rapid confirmation of influenza A H5N1 virus in clinical samples

Author: S. Corden, Guy Vernet, Pierre van Aarle, Rongbao Gao, Jean-Noel Telles, Catherine Moore, and Yue-Long Shu
Subjects: Influenza A (H5N1) Virus, Neuraminidase, Hemagglutinin Glycoproteins, Influenza Virus, Biology, medicine.disease_cause, Sensitivity and Specificity, Birds, Virology, Influenza, Human, medicine, Animals, Humans, H5N1 virus, Self-Sustained Sequence Replication, Influenza A Virus, H5N1 Subtype, Reverse Transcriptase Polymerase Chain Reaction, virus diseases, Influenza a, Influenza A/H5N1, Low pathogenic, NASBA, Influenza A virus subtype H5N1, Virus type, Influenza in Birds, Reagent Kits, Diagnostic, Sequence Alignment
Abstract: A real-time NASBA assay for the specific confirmation of influenza A H5N1 infection was developed and evaluated using proficiency panels distributed to the UK influenza network of laboratories and clinical samples received through the Chinese National Influenza Centre in Beijing. The aim of the proficiency panels was to determine the sensitivity and specificity of the assay on a range of influenza virus types and subtypes including different clades of influenza A H5 viruses. The assay was then evaluated using 19 clinical samples obtained from seven confirmed human cases of influenza A H5N1 infection in China. The assay was shown to have a level of sensitivity of 0.01 TCID50 and 10 copies/μl using RNA transcripts of the A/VietNam/1194/2004 H5N1 virus. During the evaluation the assay successfully detected H5N1 viruses known to infect humans from clades 1, 2.1, 2.2 and 2.3 as well as low pathogenic H5N3 avian influenza viruses. The clinical utility of the real-time NASBA assay was proven on a range of clinical samples from patients with confirmed H5N1 infection collected during 2005 and 2006. The real-time NASBA assay was demonstrated to be sensitive and rapid allowing for same day confirmation of a H5N1 infection direct from clinical samples.
Published: 2010
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42. Human rhinovirus C infections mirror those of human rhinovirus A in children with community-acquired pneumonia

Author: Kunling Shen, Guy Vernet, Jun Liu, Zichun Xiang, Richard Gonzalez, Gláucia Paranhos-Baccalà, Zhengde Xie, Lan Chen, Jianwei Wang, Lili Ren, Yan Xiao, Jing Zhang, Chunyan Liu, and Qi Jin
Subjects: Male, China, medicine.medical_specialty, Rhinovirus, Etiology, Community-acquired pneumonia, Adolescent, Molecular Sequence Data, Pneumonia, Viral, Prevalence, Clinical manifestation, medicine.disease_cause, Article, stomatognathic system, Nasopharynx, Virology, Internal medicine, Epidemiology, otorhinolaryngologic diseases, Humans, Medicine, Child, Children, Picornaviridae Infections, Reverse Transcriptase Polymerase Chain Reaction, business.industry, Respiratory disease, Infant, virus diseases, Sequence Analysis, DNA, medicine.disease, Community-Acquired Infections, Hospitalization, Pneumonia, Infectious Diseases, El Niño, Child, Preschool, Immunology, RNA, Viral, Female, business, circulatory and respiratory physiology
Abstract: Background Human rhinoviruses (HRVs) are among the most common causes of community-acquired pneumonia (CAP) in children. However, the differential roles of the three HRV species HRV-A, HRV-B, and HRV-C in pediatric CAP are not fully understood. Objective To determine the distribution of HRV species and their roles in children hospitalized with CAP in Beijing, China. Study design Nasopharyngeal aspirates were collected between April 2007 and March 2008 from 554 children with a primary diagnosis of CAP. HRVs in the clinical samples were detected by RT-PCR and by sequencing. Infections with other respiratory viruses were identified by PCR. Results HRVs were detected in 99 patients (17.87%). Among these patients, 51.52% tested positive for HRV-A, 38.38% for HRV-C, and 10.10% for HRV-B. HRVs were detected throughout the study period. The monthly distribution of HRV infections varied with HRV species. Median age, gender, symptoms, severity, and duration of hospitalization for single HRV-C infections were similar to those observed for single HRV-A infections. Co-infections with other respiratory viruses were detected in 57.58% of the HRV-positive children. HRV/RSV dual infections were correlated with a higher frequency of shortness of breath (HRV-A group, P2tail = 0.01; HRV-C group, P2tail = 0.015) and lower median ages (HRV-A group, P2tail = 0.049; HRV-C group, P2tail = 0.009). Conclusion Our study shows that HRV-C strains circulate at a prevalence intermediate between HRV-A and HRV-B. The severity of clinical manifestations for HRV-C is comparable to that for HRV-A in children with CAP. These findings point to an important role of both HRV-A and HRV-C in pediatric CAP.
Published: 2010
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43. Saffold Cardioviruses of 3 Lineages in Children with Respiratory Tract Infections, Beijing, China

Author: Lan Chen, Qingqing Yang, Yan Xiao, Kunling Shen, Guy Vernet, Jianwei Wang, Lili Ren, Qi Jin, Chunyan Liu, Zhengde Xie, Richard Gonzalez, Li Yongjun, and Gláucia Paranhos-Baccalà
Subjects: Microbiology (medical), Male, Pathology, medicine.medical_specialty, China, Adolescent, Epidemiology, lcsh:Medicine, Disease, respiratory tract infections, lcsh:Infectious and parasitic diseases, Beijing, medicine, Humans, viruses, lcsh:RC109-216, Respiratory system, Child, Pathogen, Phylogeny, Cardiovirus, pediatric patients, Respiratory tract infections, biology, Transmission (medicine), lcsh:R, Saffold virus, Infant, dispatch, biology.organism_classification, Virology, Infectious Diseases, Saffold cardiovirus, Child, Preschool, Female
Abstract: To clarify the potential for respiratory transmission of Saffold cardiovirus (SAFV) and characterize the pathogen, we analyzed respiratory specimens from 1,558 pediatric patients in Beijing. We detected SAFV in 7 (0.5%) patients and identified lineages 1–3. However, because 3 patients had co-infections, we could not definitively say SAFV caused disease.
Published: 2010

44. Human rhinoviruses in Chinese adults with acute respiratory tract infection

Author: Zhong Wang, Yan Xiao, Zichun Xiang, Qi Jin, Taisheng Li, Jianwei Wang, Lan Chen, Richard Gonzalez, Gláucia Paranhos-Baccalà, and Guy Vernet
Subjects: Microbiology (medical), myalgia, Adult, Male, medicine.medical_specialty, China, Adolescent, Genotype, Rhinovirus, Population, Nose, Human rhinovirus, Acute respiratory tract infections, Article, Young Adult, stomatognathic system, Throat, Internal medicine, Virology, Prevalence, otorhinolaryngologic diseases, Medicine, Adults, Humans, Respiratory system, education, Acute respiratory tract infection, Respiratory Tract Infections, Aged, Aged, 80 and over, education.field_of_study, Picornaviridae Infections, Respiratory tract infections, business.industry, Reverse Transcriptase Polymerase Chain Reaction, Respiratory disease, Chinese adults, virus diseases, Middle Aged, medicine.disease, Infectious Diseases, medicine.anatomical_structure, Immunology, Pharynx, RNA, Viral, Female, medicine.symptom, business, circulatory and respiratory physiology
Abstract: Summary Objective To evaluate the roles of human rhinoviruses (HRVs) in acute respiratory tract infections (ARTIs) in Chinese adults and determine the association between species of HRV and clinical presentations. Methods RT-PCR methods were used to detect HRVs in throat and nasal swabs collected from 6104 adult patients with ARTIs from December 2005 to April 2008 in Beijing, China. Results HRV strains were detected in 271 ARTIs cases, 65% of which tested positive for HRV-A, 25% for HRV-B, and 10% for HRV-C. Aside from fever, pharyngeal congestion and headache were the most common clinical symptoms observed in the HRVs infected patients. HRV-A infected patients had a higher percentage of upper respiratory symptoms than patients infected by the two other HRV species. Systemic symptoms such as chilliness and myalgia were more frequent in people infected by HRV-B. The three HRV species exhibited unique infection timing when analyzed monthly. Conclusion HRV-C can be detected in adult patients with acute upper respiratory tract infections, but is not the predominant species in this population.
Published: 2010

45. Prevalence of human respiratory viruses in adults with acute respiratory tract infections in Beijing, 2005–2007

Author: Qingqing Yang, Richard Gonzalez, Guy Vernet, Taisheng Li, Wei Wang, Qi Jin, Yan Xiao, Lili Ren, Y. Zhang, Hongli Zhou, J. Chen, Li Yongjun, Gláucia Paranhos-Baccalà, Lan Chen, Yaying Wang, Zichun Xiang, Zhong Wang, Jian Li, Jianwei Wang, Jing Zhang, and X. Meng
Subjects: Male, Microbiology (medical), China, Adolescent, viruses, Respiratory System, Biology, Polymerase Chain Reaction, Acute respiratory tract infections, Article, Virus, Young Adult, Age Distribution, stomatognathic system, Human metapneumovirus, Prevalence, medicine, Humans, Respiratory system, Respiratory Tract Infections, Aged, Aged, 80 and over, Respiratory tract infections, adult, Respiratory disease, virus diseases, General Medicine, Middle Aged, medicine.disease, biology.organism_classification, Virology, molecular detection, Infectious Diseases, medicine.anatomical_structure, Virus Diseases, respiratory virus infection, Acute Disease, Viruses, Immunology, Respiratory virus, Female, epidemiology, Seasons, Viral disease, Respiratory tract
Abstract: To determine the aetiological role and epidemiological profile of common respiratory viruses in adults with acute respiratory tract infections (ARTIs), a 2-year study was conducted in Beijing, China, from May 2005 to July 2007. Nose and throat swab samples from 5808 ARTI patients were analysed by PCR methods for common respiratory viruses, including influenza viruses (IFVs) A, B, and C, parainfluenza viruses (PIVs) 1–4, enteroviruses (EVs), human rhinoviruses (HRVs), respiratory syncytial virus (RSV), human metapneumovirus (HMPV), human coronaviruses (HCoVs) OC43, 229E, NL63, and HKU1, and adenoviruses (ADVs). Viral pathogens were detected in 34.6% of patient samples, and 1.6% of the patients tested positive for more than one virus. IFVs (19.3%) were the dominant agents detected, followed by HRVs (6.5%), PIVs (4.3%), EVs (3.2%), and HCoVs (1.1%). ADVs, RSV and HMPV were also detected (
Published: 2009
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46. The Impact of Dual Viral Infection in Infants Admitted to a Pediatric Intensive Care Unit Associated with Severe Bronchiolitis

Author: Audrey Bagnaud, Gláucia Paranhos-Baccalà, Geneviève Billaud, Etienne Javouhey, Daniel Floret, Guy Vernet, Florence Komurian-Pradel, Nathalie Richard, Bruno Lina, Magali Perret, and Alain Rajoharison
Subjects: Male, Microbiology (medical), Pediatrics, medicine.medical_specialty, Comorbidity, Intensive Care Units, Pediatric, law.invention, law, Intensive care, medicine, Humans, Risk factor, Intensive care medicine, Pediatric intensive care unit, business.industry, Infant, medicine.disease, Intensive care unit, Hospitalization, Natural history, Infectious Diseases, Virus Diseases, Bronchiolitis, Multivariate Analysis, Viruses, Pediatrics, Perinatology and Child Health, Pharynx, Female, Viral disease, Nasal Cavity, business
Abstract: Bronchiolitis is a major cause of morbidity and mortality in early childhood worldwide. The presence of more than one pathogen may influence the natural history of acute bronchiolitis in infants.To investigate the relevance of dual viral infection in infants with severe bronchiolitis hospitalized in a short-term unit compared with those in a pediatric intensive care unit (PICU).One hundred eighty infants1 year old hospitalized with bronchiolitis in a short-term unit (n = 92) or admitted to the PICU (n = 88) during 2 consecutive winter seasons 2003/2004 and 2004/2005 were evaluated. Molecular biology and standard methods were used to diagnose human respiratory viruses in nasal/throat swabs and nasal aspirates. Clinical data related to host factors and viral prevalence were compared among infants requiring or not PICU support.A viral agent was identified in 96.1% of infants with bronchiolitis. Respiratory syncytial virus (70.6% and 73.6%, respectively in the short-term unit and PICU) and rhinovirus (18.5% and 25.3%, respectively in the short-term unit and PICU) were the main detected respiratory viruses in infants hospitalized in both units. No significant difference in viral prevalence was observed between the populations studied. From multivariate analysis, infants with coinfections were 2.7 times (95% CI: 1.2-6.2) more at risk for PICU admission than those with a single infection. Respiratory syncytial virus and rhinovirus were the viruses most frequently identified in mixed infections in infants hospitalized with bronchiolitis.Dual viral infection is a relevant risk factor for the admission of infants with severe bronchiolitis to the PICU.
Published: 2008
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47. Comparison of reverse hybridization, microarray, and sequence analysis for genotyping hepatitis B virus

Author: Nathalie Tran, Suzan D. Pas, Robert A. de Man, Guy Vernet, Chantal Burghoorn-Maas, Hubert G. M. Niesters, Virology, and Gastroenterology & Hepatology
Subjects: Microbiology (medical), Hepatitis B virus, Genotype, Biology, medicine.disease_cause, Sensitivity and Specificity, Orthohepadnavirus, SDG 3 - Good Health and Well-being, Virology, medicine, Humans, Typing, Viral, Diagnostic, Genotyping, Phylogeny, Oligonucleotide Array Sequence Analysis, Nucleic Acid Hybridization, virus diseases, Sequence Analysis, DNA, DNA, Hepatitis B, biology.organism_classification, medicine.disease, Molecular biology, Hepadnaviridae, DNA, Viral, Reagent Kits, Reagent Kits, Diagnostic, Viral load, Sequence Analysis
Abstract: Hepatitis B virus (HBV) genotyping has become important in epidemiological and clinical diagnoses, given the relationship between the viral genotype and the progression of disease or the appearance of antiviral resistance. Since genotyping by sequence and phylogenetic analyses is not convenient in the clinical setting, we evaluated InnoLipa HBV genotyping (Innogenetics, Belgium) and an HBV DNA-Chip (bioMerieux, France) prototype assay and compared their sequencing of the gold standard S gene, using a cohort of 275 individual patient samples. All but two samples, belonging to distant and individual subgroups within a single genotype, were detected by InnoLipa HBV assay. Four samples with dual infections belonging to genotypes A and G were identified only by InnoLipa HBV assay. Using an HBV DNA-Chip assay, one sample could not be amplified due to a low viral load. Four samples were identified as genotype C and two as genotype D by sequencing but were classified as genotype A (two samples) and D (two samples) and as A (one sample) and G (one sample) by the DNA-Chip assay. In conclusion, the InnoLipa HBV genotyping strip assay detected dual infections and was an easy and quick tool for genotyping, with a sensitivity of 99.3% and a specificity of 100% compared to sequence analysis. HBV DNA-Chip assay showed a sensitivity and specificity of 97.5 and 97.8%, respectively.
Published: 2008

48. Molecular characteristics of Hepatitis B and chronic liver disease in a cohort of HB carriers from Bamako, Mali

Author: Marlin D. Friesen, Guy Vernet, Stephanie Villar, Fatou Traoré, John D. Groopman, Emmanuelle Gormally, Moussa Y Maiga, Pierre Hainaut, and Souleymane Diallo
Subjects: Adult, Liver Cirrhosis, Male, Hepatitis B virus, medicine.medical_specialty, Pathology, Adolescent, Genotype, DNA Mutational Analysis, TP53 R249S mutation, Mali, medicine.disease_cause, Chronic liver disease, Gastroenterology, Young Adult, Liver disease, Aflatoxins, Internal medicine, Chronic carriage, HBV, medicine, Humans, Viral load, Aged, Hepatitis, Hepatitis B Surface Antigens, business.industry, FibroTest, Middle Aged, Hepatitis B, Genes, p53, medicine.disease, Infectious Diseases, Carrier State, Mutation, Female, Fibrotest/Actitest, business, Research Article
Abstract: Background Hepatitis B (HB) infection is common in Mali. However, there is little information on molecular and biochemical characteristics of HB carriers. Methods A group of 1466 adult volunteers was recruited in the district of Bamako. Confirmed HB carriers were tested for HB viral load by quantitative PCR and HBV was genotyped by sequencing of HBS. Fibrosis and hepatitis activity were measured using the Fibrotest-Actitest. A mutation of TP53 at codon 249 (R249S), specific for exposure to aflatoxin, was detected in cell-free DNA extracted from plasma. Results Overall, 276 subjects were HBsAg-positive (18.8%). Among 152 subjects tested for HBV load, 49 (32.2%) had over 104 copies/mL and 16 (10.5%) had levels below the limit of detection. The E genotype was found in 91.1% of carriers. Fibrotest scores ≥ F2 were observed in 52 subjects (35.4%). Actitest scores ≥ A2 were detected in 15 subjects (10.2%) and were correlated with Fibrotest scores (p = 0.0006). Among 105 subjects tested, 60% had detectable levels of R249S copies (>40 copies/mL plasma). Conclusion Chronic HB carriage in adults in Bamako district is well over epidemic threshold. About 1/3 of carriers have moderate to severe liver fibrosis and 60% have detectable aflatoxin-related TP53 R249S mutation. These results support introduction of anti-HB therapies to reduce the progression towards severe liver disease.
Published: 2015
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49. Diagnostic values of the QuantiFERON-TB Gold In-tube assay carried out in China for diagnosing pulmonary tuberculosis

Author: Yanlin Zhao, Guy Vernet, Delia Goletti, Fabin Li, Christophe Longuet, Philippe H. Lagrange, Xiaomen Wang, and Hui Xia
Subjects: Adult, Male, China, medicine.medical_specialty, animal structures, Tuberculosis, QUANTIFERON-TB GOLD, Human immunodeficiency virus (HIV), Tuberculin, lcsh:Medicine, medicine.disease_cause, Sensitivity and Specificity, Young Adult, Pulmonary tuberculosis, Internal medicine, Humans, Medicine, lcsh:Science, Tuberculosis, Pulmonary, Multidisciplinary, integumentary system, business.industry, lcsh:R, Skin test, Middle Aged, medicine.disease, bacterial infections and mycoses, Case-Control Studies, Immunology, Female, lcsh:Q, Reagent Kits, Diagnostic, business, Research Article
Abstract: Background Interferon-release assays (IGRAs) for diagnosing active pulmonary tuberculosis (PTB) are not yet fully validated, particularly in high TB-endemic areas as the People's Republic of China (PRC). The aim of this report was to assess the performance of the QuantiFERON-TB Gold In-tube (QFT-GIT) and tuberculin skin test (TST), in addition to microbiological results, as contributors for diagnosing active PTB in the PRC. Methods/Principal Findings A total of 300 PTB patients, 41 disease controls (DC) and 59 healthy community controls (HCC) were included prospectively between May 2010 and April 2011 from two provinces of the PRC (Heilongjiang and Zhejiang). The QFT-GIT and TST yielded an overall sensitivity for active TB of 80.9% and 86.2%, and a specificity of 36.6% and 26.8%, respectively. The province of origin and smear microscopy status did not significantly impact the diagnostic values for PTB. However, using the TST with a 10 mm cut-off point, a significantly higher proportion of LTBI was observed in the DC than the HCC (p=0.01). Discordant results between the QFT-GIT and TST were found among 1/3 of the PTB, HCC and DC. Two-thirds of the individuals presented TST-positive/QFT-GIT-negative discordant results. The TST-negative/QFT-GIT-positive result was not associated with age or bacillary load. Cumulative QFT-GIT and TST positive results increased the overall sensitivity (95.9%), but it was associated with a dramatic decrease of the overall specificity (24.8%) leading to a suboptimal PPV (80.1%) and a low NPV (61.1%). Conclusions/Significance The usefulness of the QFT-GIT to diagnose active TB in high TB-endemic countries remains doubtful because like the TST, the QFT-GIT cannot distinguish between LTBI and active TB. Used as single stand-alone tests, both the QFT-GIT and TST have very limited roles in the diagnosis of active PTB. However, the combined use of SM, the TST and QFT-GIT may allow for the exclusion of ATB.
Published: 2015

50. Molecular epidemiology of hepatitis B virus in Dakar, Sénégal

Author: Papa Saliou Mb, Benoit Chevalier, Jean-Christophe Plantier, Jean-Marie Sire, François Simon, Nathalie Tran, Muriel Vray, Fatou Fall, Guy Vernet, and Jean-Marc Debonne
Subjects: Adult, Male, Hepatitis B virus, Genotype, Population, Blood Donors, Hepacivirus, medicine.disease_cause, Polymerase Chain Reaction, Hepatitis B, Chronic, Orthohepadnavirus, Virology, medicine, Humans, Point Mutation, Hepatitis B e Antigens, education, Oligonucleotide Array Sequence Analysis, Molecular Epidemiology, education.field_of_study, Hepatitis B Surface Antigens, biology, Molecular epidemiology, Alanine Transaminase, Middle Aged, Viral Load, Hepatitis B, biology.organism_classification, medicine.disease, Hepatitis C, Hepatitis D, Senegal, Infectious Diseases, Hepadnaviridae, Codon, Nonsense, DNA, Viral, Immunology, Female, Viral load
Abstract: Using DNA chip technology and real-time quantitative PCR, molecular profile of HBV strains infecting blood donors and patients in Dakar, Senegal was studied. All HBsAg-positive blood donors (n = 175) and all patients who presented with chronic hepatitis B (n = 29) between 1st June 2003 and 31st July 2003 were studied. One patient, a blood donor, was coinfected by HCV, and nine patients had anti-HDV antibodies. Few persons in either group were HBeAg-positive. Viral load values were relatively low but correlated with biochemical abnormalities. Patients were infected mainly by genotype E (72%). Patients infected by genotype A (28%) tended to be younger than other patients. There was no significant difference between the blood donors and the patients with hepatitis B as regards virological markers, including viral load, when the HBV genotype was taken into account. The BCP A1762T and G1764A mutations were found in four patients and one patient, respectively; the two mutations were never found in the same patient. The W28* mutation at position 1896 of the core was detected in 19 of the 32 genotyped patients, 18 (83%) of whom had genotype E infection. ALT levels were not influenced by HBV mutations. This study shows a low frequency of clinical signs in HBsAg-positive blood donors, a relatively low level of viral replication, and a high frequency of pre-core mutants in this West African population. These results underline the importance of molecular characterization of HBV infection as specific treatments become available in this region. J. Med. Virol. 78:329–334, 2006. © 2006 Wiley-Liss, Inc.
Published: 2006
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