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1. Genomic surveillance reveals a dengue 2 virus epidemic lineage with a marked decrease in sensitivity to Mosnodenvir

Author

Hawa Sophia Bouzidi, Selin Sen, Géraldine Piorkowski, Laura Pezzi, Nazli Ayhan, Albin Fontaine, Thomas Canivez, Manon Geulen, Rayane Amaral, Gilda Grard, Guillaume André Durand, Xavier de Lamballerie, Franck Touret, and Raphaëlle Klitting

Subjects
Science
Abstract

Abstract Dengue fever is the most important arbovirosis for public health, with more than 5 million cases worldwide in 2023. Mosnodenvir is the first anti-dengue compound with very high preclinical pan-serotype activity, currently undergoing phase 2 clinical evaluation. Here, by analyzing dengue virus (DENV) genomes from the 2023-2024 epidemic in the French Caribbean Islands, we show that they all exhibit mutation NS4B:V91A, previously associated with a marked decrease in sensitivity to mosnodenvir in vitro. Using antiviral activity tests on four clinical and reverse-genetic strains, we confirm a marked decrease in mosnodenvir sensitivity for DENV-2 ( > 1000 fold). Finally, combining phylogenetic analysis and experimental testing for resistance, we find that virus lineages with low sensitivity to mosnodenvir due to the V91A mutation likely emerged multiple times over the last 30 years in DENV-2 and DENV-3. These results call for increased genomic surveillance, in particular to track lineages with resistance mutations. These efforts should allow to better assess the activity profile of DENV treatments in development against circulating strains.

Published
2024
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2. Analytical and clinical evaluation of a duplex RT-qPCR assay for the detection and identification of o’nyong-nyong and chikungunya virus

Author

Konrad M. Wesselmann, Lea Luciani, Laurence Thirion, Xavier de Lamballerie, Remi Charrel, and Laura Pezzi

Subjects
Alphavirus, Togaviridae, arbovirus, chikungunya virus, o’nyong-nyong virus, diagnostics, Infectious and parasitic diseases, RC109-216, Microbiology, QR1-502
Abstract

The mosquito-borne alphavirus o’nyong-nyong virus (ONNV) has proven its potential to cause major human outbreaks. On the African continent, ONNV causes unspecific febrile illness and co-circulates with the close relative chikungunya virus (CHIKV). The true scale of ONNV burden is poorly understood in Africa, because of the scarce availability of molecular in-house and commercial assays, strong cross-reactivity between ONNV and CHIKV in serological assays and a lack of surveillance. We designed a new RT-qPCR assay targeting the E1 gene for the detection of ONNV that can be used in monoplex or in duplex format, combined with a previously published CHIKV monoplex assay targeting the nsp1. The lower limit of detection with 95% positivity rate was determined to be

Published
2024
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3. Broad-spectrum dengue virus detection using the commercial RealStar dengue RT-PCR kit 3.0 (Altona) and an in-house combined real-time RT-PCR assay

Author

Léa Luciani, Pierre Combe, Franck Touret, Céline Gazin, Raphaëlle Klitting, Laura Pezzi, Laurence Thirion, Rémi Charrel, Gilda Grard, Xavier de Lamballerie, and Antoine Nougairède

Subjects
RT-PCR, Dengue virus, Diagnostic, Science (General), Q1-390, Social sciences (General), H1-99
Abstract

In endemic areas, the genetic diversity among co-circulating dengue virus (DENV) strains is considerable and new, highly divergent strains are identified on a regular basis. It is thus critical to ensure that molecular diagnostic tools effectively detect virus genomes even in case of important genetic variation. Here, we tested both the pan-DENV detection capacity and the limit of detection of two real-time RT-PCR assays: (i) the commercial RealStar Altona 3.0 system and (ii) a laboratory developed test (LDT) combining two RT-PCR systems in a single reaction tube (DenAllDUO). We used a panel of DENV strains representative of the genetic diversity within DENV species, combined with three in vitro transcribed RNAs as surrogates for unavailable strains corresponding to recently discovered strains with substantial genetic divergence: DENV serotype 1 (DENV-1) Brun2014, DENV-2 QML22 and DENV-4 DKE121. Both systems (i) targeted the genome 3′ untranslated region, (ii) displayed a broad detection spectrum, encompassing most of DENV species diversity, and (iii) detected the three aforementioned divergent strains. DenAllDUO detected all the strains tested, whereas the RealStar system failed to detect strains from DENV-4 genotype III. Altogether, our findings support the value of these two RT-PCR systems as part of the Dengue diagnostic arsenal.

Published
2024
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4. Assessing vulnerability for future Zika virus outbreaks using seroprevalence data and environmental suitability maps.

Author

Yannik Roell, Laura Pezzi, Anyela Lozano-Parra, Daniel Olson, Jane Messina, Talia Quandelacy, Jan Felix Drexler, Oliver Brady, Morteza Karimzadeh, and Thomas Jaenisch

Subjects
Arctic medicine. Tropical medicine, RC955-962, Public aspects of medicine, RA1-1270
Abstract

The 2015-17 Zika virus (ZIKV) epidemic in the Americas subsided faster than expected and evolving population immunity was postulated to be the main reason. Herd immunization is suggested to occur around 60-70% seroprevalence, depending on demographic density and climate suitability. However, herd immunity was only documented for a few cities in South America, meaning a substantial portion of the population might still be vulnerable to a future Zika virus outbreak. The aim of our study was to determine the vulnerability of populations to ZIKV by comparing the environmental suitability of ZIKV transmission to the observed seroprevalence, based on published studies. Using a systematic search, we collected seroprevalence and geospatial data for 119 unique locations from 37 studies. Extracting the environmental suitability at each location and converting to a hypothetical expected seroprevalence, we were able to determine the discrepancy between observed and expected. This discrepancy is an indicator of vulnerability and divided into three categories: high risk, low risk, and very low risk. The vulnerability was used to evaluate the level of risk that each location still has for a ZIKV outbreak to occur. Of the 119 unique locations, 69 locations (58%) fell within the high risk category, 47 locations (39%) fell within the low risk category, and 3 locations (3%) fell within the very low risk category. The considerable heterogeneity between environmental suitability and seroprevalence potentially leaves a large population vulnerable to future infection. Vulnerability seems to be especially pronounced at the fringes of the environmental suitability for ZIKV (e.g. Sao Paulo, Brazil). The discrepancies between observed and expected seroprevalence raise the question: "why did the ZIKV epidemic stop with large populations unaffected?". This lack of understanding also highlights that future ZIKV outbreaks currently cannot be predicted with confidence.

Published
2024
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5. Current Antithrombotic Treatments for Cardiovascular Diseases: A Comprehensive Review

Author

Kristian Galanti, Mario Di Marino, Davide Mansour, Sabrina Testa, Davide Rossi, Claudio Scollo, Roberta Magnano, Laura Pezzi, Alberto D'Alleva, Daniele Forlani, Piergiusto Vitulli, Leonardo Paloscia, Fabrizio Ricci, Giulia Renda, Sabina Gallina, and Massimo Di Marco

Subjects
antithrombotic treatments, anticoagulation therapy, antiplatelet therapy, ischemic risk, bleeding risk, risk-to-benefit ratio, cardiovascular diseases, Diseases of the circulatory (Cardiovascular) system, RC666-701
Abstract

Antithrombotic therapies (ATT) play a pivotal role in the management of cardiovascular diseases, aiming to prevent ischemic events while maintaining a delicate balance with the patient’s bleeding risk. Typically, ATT can be classified into antiplatelet and anticoagulant therapies. Their application spans a broad spectrum of cardiovascular conditions, ranging from ischemic heart disease to atrial fibrillation, encompassing venous thromboembolisms and innovative structural interventional cardiology procedures. The global burden of cardiovascular diseases is steadily increasing, often giving rise to overlapping clinical presentations. Accordingly, the adoption of combined pharmacological approaches becomes imperative, potentially disrupting the delicate equilibrium between ischemic and bleeding risk, thus leading to nuanced pharmacotherapeutic pathways. In this context, contemporary investigations strive to identify a convergence point that optimizes the duration of medical therapy while addressing the need for antithrombotic effects, especially in the context of ischemic heart disease. This review aims to comprehensively revisit the main antithrombotic strategies in cardiovascular diseases, with the intention of enhancing a systematic approach which is key for the effective clinical management of these patients. Also, the review will examine the most impactful studies that have established the groundwork for current scientific evidence, with acknowledgement of special populations. Finally, we will cast a gaze into the future of this dynamic and evolving research field, exploring forthcoming perspectives and advancements.

Published
2024
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6. Molecular detection of parapoxvirus in Ixodidae ticks collected from cattle in Corsica, France

Author

Vincent Cicculli, Nazli Ayhan, Léa Luciani, Laura Pezzi, Apolline Maitre, Dorine Decarreaux, Xavier de Lamballerie, Jean‐Christophe Paoli, Laurence Vial, Remi Charrel, and Alessandra Falchi

Subjects
cattle, epidemiology, ticks, zoonoses, Veterinary medicine, SF600-1100
Abstract

Abstract Background Several viruses belonging to the family Poxviridae can cause infections in humans and animals. In Corsica, livestock farming (sheep, goats, pigs, and cattle) is mainly mixed, leading to important interactions between livestock, wildlife, and human populations. This could facilitate the circulation of zoonotic diseases, and makes Corsica a good example for studies of tick‐borne diseases. Objectives To gain understanding on the circulation of poxviruses in Corsica, we investigated their presence in tick species collected from cattle, sheep, horses, and wild boar, and characterized them through molecular techniques. Methods Ticks were tested using specific primers targeting conserved regions of sequences corresponding to two genera: parapoxvirus and orthopoxvirus. Results A total of 3555 ticks were collected from 1549 different animals (687 cattle, 538 horses, 106 sheep, and 218 wild boars). They were tested for the presence of parapoxvirus DNA on one hand and orthopoxvirus DNA on the other hand using Pangeneric real‐time TaqMan assays. Orthopoxvirus DNA was detected in none of the 3555 ticks. Parapoxvirus DNA was detected in 6.6% (36/544) of ticks collected from 23 cows from 20 farms. The remaining 3011 ticks collected from horses, wild boars, and sheep were negative. The infection rate in cow ticks was 8.0% (12/148) in 2018 and 6.0% (24/396) in 2019 (p = 0.57). Parapoxvirus DNA was detected in 8.5% (5/59) of Hyalomma scupense pools, 8.2% (15/183) of Hyalomma marginatum pools, and 6.7% (16/240) of Rhipicephalus bursa pools (p = 0.73). We successfully amplified and sequenced 19.4% (7/36) of the positive samples which all corresponded to pseudocowpox virus. Conclusions Obviously, further studies are needed to investigate the zoonotic potential of pseudocowpox virus and its importance for animals and public health.

Published
2022
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7. Specimen sharing for epidemic preparedness: Building a virtual biorepository system from local governance to global partnerships.

Author

Judith Giri, Laura Pezzi, Rodrigo Cachay, Rosa Margarita Gèlvez Ramirez, Adriana Tami, Sarah Bethencourt, Anyela Lozano, José Eduardo Gotuzzo Herencia, Julia Poje, Thomas Jaenisch, and May Chu

Subjects
Public aspects of medicine, RA1-1270
Abstract

We present a framework for a federated, virtual biorepository system (VBS) with locally collected and managed specimens, as a 'global public good' model based on principles of equitable access and benefit sharing. The VBS is intended to facilitate timely access to biological specimens and associated data for outbreak-prone infectious diseases to accelerate the development and evaluation of diagnostics, assess vaccine efficacy, and to support surveillance and research needs. The VBS is aimed to be aligned with the WHO BioHub and other specimen sharing efforts as a force multiplier to meet the needs of strengthening global tools for countering epidemics. The purpose of our initial research is to lay the basis of the collaboration, management and principles of equitable sharing focused on low- and middle-income country partners. Here we report on surveys and interviews undertaken with biorepository-interested parties to better understand needs and barriers for specimen access and share examples from the ZIKAlliance partnership on the governance and operations of locally organized biorepositories.

Published
2023
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8. Model-based assessment of Chikungunya and O’nyong-nyong virus circulation in Mali in a serological cross-reactivity context

Author

Nathanaël Hozé, Issa Diarra, Abdoul Karim Sangaré, Boris Pastorino, Laura Pezzi, Bourèma Kouriba, Issaka Sagara, Abdoulaye Dabo, Abdoulaye Djimdé, Mahamadou Ali Thera, Ogobara K. Doumbo, Xavier de Lamballerie, and Simon Cauchemez

Subjects
Science
Abstract

Abstract Serological surveys are essential to quantify immunity in a population but serological cross-reactivity often impairs estimates of the seroprevalence. Here, we show that modeling helps addressing this key challenge by considering the important cross-reactivity between Chikungunya (CHIKV) and O’nyong-nyong virus (ONNV) as a case study. We develop a statistical model to assess the epidemiology of these viruses in Mali. We additionally calibrate the model with paired virus neutralization titers in the French West Indies, a region with known CHIKV circulation but no ONNV. In Mali, the model estimate of ONNV and CHIKV prevalence is 30% and 13%, respectively, versus 27% and 2% in non-adjusted estimates. While a CHIKV infection induces an ONNV response in 80% of cases, an ONNV infection leads to a cross-reactive CHIKV response in only 22% of cases. Our study shows the importance of conducting serological assays on multiple cross-reactive pathogens to estimate levels of virus circulation.

Published
2021
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11. Detection and Isolation of Sindbis Virus from Field Collected Mosquitoes in Timimoun, Algeria

Author

Nazli Ayhan, Aissam Hachid, Laurence Thirion, Kamel Eddine Benallal, Laura Pezzi, Fayez Ahmed Khardine, Chahrazed Benbetka, Sihem Benbetka, Zoubir Harrat, and Remi Charrel

Subjects
Sindbis virus, alphavirus, Togaviridae, Culex pipiens, Culex perexiguus, arbovirus, Microbiology, QR1-502
Abstract

Sindbis virus (SINV) is a zoonotic alphavirus (family Togaviridae, genus Alphavirus) that causes human diseases in Africa, Europe, Asia, and Australia. Occasionally, SINV outbreaks were reported in South Africa and northern Europe. Birds are the main amplifying hosts of SINV, while mosquitoes play the role of the primary vector. Culex mosquitoes were collected in Algeria and subsequently tested for SINV. SINV RNA was detected in 10 pools out of 40, from a total of 922 mosquitoes tested. A strain of SINV was isolated from a pool displaying high viral load. Whole-genome sequencing and phylogenetic analysis showed that the SINV Algeria isolate was most closely related to a Kenyan strain. This was the first record of SINV in Algeria and more broadly in northwestern Africa, which can be a potential risk for human health in the circulating area. Further studies are needed to measure the impact on public health through seroprevalence studies in Algeria.

Published
2022
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12. Brainstem Correlates of a Cold Pressor Test Measured by Ultra-High Field fMRI

Author

Mariëlle C. Hendriks-Balk, Fatma Megdiche, Laura Pezzi, Olivier Reynaud, Sandra Da Costa, Domenica Bueti, Dimitri Van De Ville, and Grégoire Wuerzner

Subjects
cold pressor test, brainstem, BOLD, fMRI, blood pressure, sympathetic nervous system, Neurosciences. Biological psychiatry. Neuropsychiatry, RC321-571
Abstract

IntroductionModern imaging techniques such as blood oxygen level-dependent (BOLD) functional magnetic resonance imaging (fMRI) allow the non-invasive and indirect measurement of brain activity. Whether changes in signal intensity can be detected in small brainstem regions during a cold pressor test (CPT) has not been explored thoroughly. The aim of this study was to measure whole brain and brainstem BOLD signal intensity changes in response to a modified CPT.MethodsBOLD fMRI was measured in healthy normotensive participants during a randomized crossover study (modified CPT vs. control test) using ultra-high field 7 Tesla MRI scanner. Data were analyzed using Statistical Parametric Mapping (SPM) in a whole-brain approach, and with a brainstem-specific analysis using the spatially unbiased infra-tentorial template (SUIT) toolbox. Blood pressure (BP) and hormonal responses (norepinephrine and epinephrine levels) were also measured. Paired t-test statistics were used to compare conditions.ResultsEleven participants (six women, mean age 28 ± 8.9 years) were analyzed. Mean arterial BP increased from 83 ± 12 mm Hg to 87 ± 12 mm Hg (p = 0.0009) during the CPT. Whole-brain analysis revealed significant activations linked to the CPT in the right supplementary motor cortex, midcingulate (bilateral) and the right anterior insular cortex. The brainstem-specific analysis showed significant activations in the dorsal medulla.ConclusionChanges in BOLD fMRI signal intensity in brainstem regions during a CPT can be detected, and show an increased response during a cold stress in healthy volunteers. Consequently, BOLD fMRI at 7T is a promising tool to explore and acquire new insights in the comprehension of neurogenic hypertension.

Published
2020
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13. High SARS-CoV-2 Prevalence among Healthcare Workers in Cochabamba, Bolivia

Author

Paola Mariela Saba Villarroel, María del Rosario Castro Soto, Verónica Undurraga, Heydi Sanz, Ana María Jaldín, Laetitia Ninove, Elif Nurtop, Laura Pezzi, Souand Mohamed Ali, Abdennour Amroun, Morgan Seston, and Xavier de Lamballerie

Subjects
seroprevalence, SARS-CoV-2, healthcare workers, Bolivia, Microbiology, QR1-502
Abstract

Healthcare workers (HCWs) are at increased risk of SARS-CoV-2 infection. The aim of the study was to estimate the SARS-CoV-2 seroprevalence among HCWs in Cochabamba, Bolivia and to determine the potential risk factors. In January 2021, a cross-sectional SARS-CoV-2 seroprevalence study was conducted in 783 volunteer clinical and non-clinical HCWs in tertiary care facilities. It was based on IgG detection using ELISA, chemiluminiscence, and seroneutralisation tests from dried blood spots. Analysis revealed a high seroprevalence (43.4%) of SARS-CoV-2 IgG antibodies. The combination of anosmia and ageusia (OR: 68.11; 95%-CI 24.83–186.80) was predictive of seropositivity. Belonging to the cleaning staff (OR: 1.94; 95%-CI 1.09–3.45), having more than two children in the same house (OR: 1.74; 95%-CI 1.12–2.71), and having been in contact with a close relative with COVID-19 (OR: 3.53; 95%-CI 2.24–5.58) were identified as risk factors for seropositivity in a multivariate analysis. A total of 47.5% of participants had received medication for COVID-19 treatment or prevention, and only ~50% of symptomatic subjects accessed PCR or antigenic testing. This study confirms a massive SARS-CoV-2 attack rate among HCWs in Cochabamba by the end of January 2021. The main risk factors identified are having a low-skilled job, living with children, and having been in contact with an infected relative in the household.

Published
2022
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14. Evaluation of a Trio Toscana Virus Real-Time RT-PCR Assay Targeting Three Genomic Regions within Nucleoprotein Gene

Author

Laurence Thirion, Laura Pezzi, Irene Pedrosa-Corral, Sara Sanbonmatsu-Gamez, Xavier De Lamballerie, Alessandra Falchi, Mercedes Perez-Ruiz, and Remi N. Charrel

Subjects
Phlebovirus, Phenuiviridae, meningitis, sand fly, Phlebotomus, arbovirus, Medicine
Abstract

Toscana virus (TOSV) can cause central nervous system infections in both residents of and travelers to Mediterranean countries. Data mining identified three real-time RT-qPCR assays for detecting TOSV RNA targeting non-overlapping regions in the nucleoprotein gene. Here, they were combined to create a multi-region assay named Trio TOSV RT-qPCR consisting of six primers and three probes. In this study, (i) we evaluated in silico the three RT-qPCR assays available in the literature for TOSV detection, (ii) we combined the three systems to create the Trio TOSV RT-qPCR, (iii) we assessed the specificity and sensitivity of the three monoplex assays versus the Trio TOSV RT-qPCR assay, and (iv) we compared the performance of the Trio TOSV RT-qPCR assay with one of the reference monoplex assays on clinical samples. In conclusion, the Trio TOSV RT-qPCR assay performs equally or better than the three monoplex assays; therefore, it provides a robust assay that can be used for both research and diagnostic purposes.

Published
2021
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15. Development and Evaluation of a duo SARS-CoV-2 RT-qPCR Assay Combining Two Assays Approved by the World Health Organization Targeting the Envelope and the RNA-Dependant RNA Polymerase (RdRp) Coding Regions

Author

Laura Pezzi, Remi N. Charrel, Laetitia Ninove, Antoine Nougairede, Gregory Molle, Bruno Coutard, Guillaume Durand, Isabelle Leparc-Goffart, Xavier de Lamballerie, and Laurence Thirion

Subjects
coronavirus, COVID-19, real-time PCR, diagnostics, diagnosis, emerging, Microbiology, QR1-502
Abstract

The recent emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) worldwide has highlighted the importance of reliable and rapid diagnostic testing to prevent and control virus circulation. Dozens of monoplex in-house RT-qPCR assays are already available; however, the development of dual-target assays is suited to avoid false-negative results caused by polymorphisms or point mutations, that can compromise the accuracy of diagnostic and screening tests. In this study, two mono-target assays recommended by WHO (E-Sarbeco (enveloppe gene, Charite University, Berlin, Germany) and RdRp-IP4 (RdRp, Institut Pasteur, Paris, France)) were selected and combined in a unique robust test; the resulting duo SARS-CoV-2 RT-qPCR assay was compared to the two parental monoplex tests. The duo SARS-CoV-2 assay performed equally, or better, in terms of sensitivity, specificity, linearity and signal intensity. We demonstrated that combining two single systems into a dual-target assay (with or without an MS2-based internal control) did not impair performances, providing a potent tool adapted for routine molecular diagnosis in clinical microbiology laboratories.

Published
2020
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16. Detection of Two Species of the Genus Parapoxvirus (Bovine Papular Stomatitis Virus and Pseudocowpox Virus) in Ticks Infesting Cattle in Burkina Faso

Author

Achille Ouedraogo, Léa Luciani, Olivier Zannou, Abel Biguezoton, Laura Pezzi, Laurence Thirion, Adrien Belem, Claude Saegerman, Rémi Charrel, and Laetitia Lempereur

Subjects
bovine papular stomatitis, pseudocowpox, virus, arbovirus, poxviridae, cattle ticks, Biology (General), QH301-705.5
Abstract

The molecular identification of arboviruses in West Africa is of particular interest, due to their zoonotic potential in a population living in close contact with livestock, and in a region where the livestock migration across borders raises the risk of diseases infection and dissemination. The aim of the study was the screening of potential circulating arboviruses and the assessment of their zoonotic implications. Therefore, ticks were collected on cattle located in three provinces of eastern Burkina Faso. Tick pools were tested using a panel of genus-specific real-time assays targeting conserved regions of parapoxvirus, orthopoxvirus, flavivirus and phlebovirus. On the 26 farms visited, a total of 663 ticks were collected. Four genera and six tick species were morphologically identified, with Amblyomma variegatum and Hyalomma spp. being the most represented species. No arboviruses were found. However, this study highlights the presence of pseudocowpox virus (8.2%) and bovine papular stomatitis virus (5.8%) among the positive tick pools. BPSV positive ticks were found in herds sharing water and pastures resources and with a history of seasonal transhumance. Therefore, common grazing and the seasonal transhumance are likely to support the transmission of the virus. This could have important health and economic impacts, especially regarding transboundary cattle movements.

Published
2020
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17. Lyophilized Matrix Containing Ready-to-Use Primers and Probe Solution for Standardization of Real-Time PCR and RT-qPCR Diagnostics in Virology

Author

Laurence Thirion, Audrey Dubot-Peres, Laura Pezzi, Iban Corcostegui, Mhammed Touinssi, Xavier de Lamballerie, and Remi N. Charrel

Subjects
freeze-drying, lyophilization, pcr, diagnosis, virus, pathogen, taqman, emerging, epidemic, Microbiology, QR1-502
Abstract

Real-time molecular techniques have become the reference methods for direct diagnosis of pathogens. The reduction of steps is a key factor in order to decrease the risk of human errors resulting in invalid series and delayed results. We describe here a process of preparation of oligonucleotide primers and hydrolysis probe in a single tube at predefined optimized concentrations that are stabilized via lyophilization (Lyoph-P&P). Lyoph-P&P was compared versus the classic protocol using extemporaneously prepared liquid reagents using (i) sensitivity study, (ii) long-term stability at 4 °C, and (iii) long-term stability at 37 °C mimicking transportation without cold chain. Two previously published molecular assays were selected for this study. They target two emerging viruses that are listed on the blueprint of the WHO as to be considered for preparedness and response actions: chikungunya virus (CHIKV) and Rift Valley fever phlebovirus (RVFV). Results of our study demonstrate that (i) Lyoph-P&P is stable for at least 4 days at 37 °C supporting shipping without the need of cold chain, (ii) Lyoph-P&P rehydrated solution is stable at +4 °C for at least two weeks, (iii) sensitivity observed with Lyoph-P&P is at least equal to, often better than, that observed with liquid formulation, (iv) validation of results observed with low-copy specimens is rendered easier by higher fluorescence level. In conclusion, Lyoph-P&P holds several advantages over extemporaneously preparer liquid formulation that merit to be considered when a novel real-time molecular assay is implemented in a laboratory in charge of routine diagnostic activity.

Published
2020
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18. Development and Evaluation of a Duo Chikungunya Virus Real-Time RT-PCR Assay Targeting Two Regions within the Genome

Author

Laurence Thirion, Laura Pezzi, Iban Corcostegui, Audrey Dubot-Pérès, Alessandra Falchi, Xavier de Lamballerie, and Remi N. Charrel

Subjects
Alphavirus, Togaviridae, emergence, arbovirus, arthropod-borne, fever, epidemic, outbreak, tropical disease, Microbiology, QR1-502
Abstract

Chikungunya virus (CHIKV) re-emerged as a globalized health threat fifteen years ago. There are dozens of RT-PCR assays published. An inventory of the latter was made, and after in silico analysis, two assays were selected for their ability to detect strains belonging to the five CHIKV genetic lineages. They were combined in order to provide a robust assay not affected by genetic point mutations and the resulting Duo CHIKV real-time RT-PCR assay was compared to the two parental single-plex tests against five strains belonging to the five genetic lineages. The Duo CHIKV assay performed equally, or better, in terms of sensitivity, specificity, linearity and signal intensity. Dual-target assays are better suited for viruses having the propensity to evolve into new variants via point mutations or major sequence deletions/insertions. Here, we demonstrated that combining two single systems into a dual-target assay did not impair sensitivity and specificity, and proved a potent diagnostic tool to face a potential emergence of CHIKV variants by newly evolving mutations.

Published
2019
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19. Limitations introduced by a low participation rate of SARS-CoV-2 seroprevalence data

Author

Olivia Pluss, Harlan Campbell, Laura Pezzi, Ivonne Morales, Yannik Roell, Talia M Quandelacy, Rahul Krishan Arora, Emily Boucher, Molly M Lamb, May Chu, Till Bärnighausen, and Thomas Jaenisch

Subjects
Epidemiology, General Medicine
Abstract

Background There has been a large influx of COVID-19 seroprevalence studies, but comparability between the seroprevalence estimates has been an issue because of heterogeneities in testing platforms and study methodology. One potential source of heterogeneity is the response or participation rate. Methods We conducted a review of participation rates (PR) in SARS-CoV-2 seroprevalence studies collected by SeroTracker and examined their effect on the validity of study conclusions. PR was calculated as the count of participants for whom the investigators had collected a valid sample, divided by the number of people invited to participate in the study. A multivariable beta generalized linear model with logit link was fitted to determine if the PR of international household and community-based seroprevalence studies was associated with the factors of interest, from 1 December 2019 to 10 March 2021. Results We identified 90 papers based on screening and were able to calculate the PR for 35 out of 90 papers (39%), with a median PR of 70% and an interquartile range of 40.92; 61% of the studies did not report PR. Conclusions Many SARS-CoV-2 seroprevalence studies do not report PR. It is unclear what the median PR rate would be had a larger portion not had limitations in reporting. Low participation rates indicate limited representativeness of results. Non-probabilistic sampling frames were associated with higher participation rates but may be less representative. Standardized definitions of participation rate and data reporting necessary for the PR calculations are essential for understanding the representativeness of seroprevalence estimates in the population of interest.

Published
2022

20. FHL1 is a major host factor for chikungunya virus infection

Author

Cécile Doyen, Thérèse Couderc, Alexis Brugier, R. Juntas-Morales, Philippe Roingeard, Rabah Ben Yaou, Julien Burlaud-Gaillard, Etienne Simon-Loriere, Mohamed Lamine Hafirassou, Sophia Rafasse, Vasiliya Kril, Pierre-Olivier Vidalain, Laura Pezzi, Gisèle Bonne, Lucie Gueneau, Xavier de Lamballerie, Marc Lecuit, Laurent Meertens, Anne Bertrand-Legout, Beate M. Kümmerer, Ali Amara, Lucie Bonnet-Madin, Thibaud Goupil, Constance Delaugerre, Athena Labeau, Monsef Benkirane, Génomes, biologie cellulaire et thérapeutiques (GenCellDi (UMR_S_944)), Collège de France (CdF (institution))-Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Hôpital Saint-Louis, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Diderot - Paris 7 (UPD7), Biologie des Infections - Biology of Infection, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institute of Virology [Bonn, Germany], University of Bonn Medical Centre, Génomique évolutive des virus à ARN - Evolutionary genomics of RNA viruses, Institut Pasteur [Paris] (IP), Morphogénèse et antigénicité du VIH et du virus des Hépatites (MAVIVH - U1259 Inserm - CHRU Tours ), Centre Hospitalier Régional Universitaire de Tours (CHRU Tours)-Université de Tours (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de génétique humaine (IGH), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Unité des Virus Emergents (UVE), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de Chimie et de Biochimie Pharmacologiques et Toxicologiques (LCBPT - UMR 8601), Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Equipe Chimie et Biologie, Modélisation et Immunologie pour la Thérapie (CBMIT), Université Paris Descartes - Paris 5 (UPD5)-Université Paris Descartes - Paris 5 (UPD5), Génomique virale et vaccination, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Centre de recherche en Myologie – U974 SU-INSERM, Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), Département de neurologie [Montpellier], Université Montpellier 1 (UM1)-Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier)-Hôpital Gui de Chauliac [CHU Montpellier], Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier)-Université de Montpellier (UM), Laboratoire de Virologie [CHU Saint-Louis], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Groupe Hospitalier Saint Louis - Lariboisière - Fernand Widal [Paris], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Service de maladies infectieuses et tropicales [Saint-Louis], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Diderot - Paris 7 (UPD7)-Groupe Hospitalier Saint Louis - Lariboisière - Fernand Widal [Paris], Service des Maladies infectieuses et tropicales [CHU Necker], CHU Necker - Enfants Malades [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), This study received funding from the French Government’s Investissement d’Avenir program, Laboratoire d’Excellence ‘Integrative Biology of Emerging Infectious Diseases’ (ANR-10-LABX-62-IBEID), the ‘Investissements d’Avenir’ program (ANR-10-IHUB-0002, ANR-15-CE15-00029 ZIKAHOST and the INCEPTION program ANR-16-CONV-0005), Institut Pasteur, Inserm and European Research Council. Viruses were provided by the Europe and Virus Archive, which has received funding from the EU Horizon 2020 research and innovation program under grant agreement 653316. The authors thank the EA7310, Laboratoire de Virologie, Université de Corse-Inserm for funding the doctoral position of L.P., ANR-10-LABX-0062,IBEID,Integrative Biology of Emerging Infectious Diseases(2010), ANR-17-CE15-0029,ZIKAHOST,LES FACTEURS DE L'HÔTE ASSOCIÉS À LA NEUROPATHOGÉNICITÉ DU VIRUS ZIKA(2017), ANR-16-CONV-0005,INCEPTION,Institut Convergences pour l'étude de l'Emergence des Pathologies au Travers des Individus et des populatiONs(2016), Institut National de la Santé et de la Recherche Médicale (INSERM)-Collège de France (CdF (institution))-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Pasteur [Paris], Université de Tours-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Hospitalier Régional Universitaire de Tours (CHRU TOURS), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Centre de Recherche en Myologie, Université Montpellier 1 (UM1)-Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier)-Hôpital Gui de Chauliac [Montpellier]-Université de Montpellier (UM), Université Paris Diderot - Paris 7 (UPD7)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Groupe Hospitalier Saint Louis - Lariboisière - Fernand Widal [Paris], Aix Marseille Université (AMU)-Institut de Recherche pour le Développement (IRD)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris], Hôpital Gui de Chauliac [Montpellier]-Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier)-Université Montpellier 1 (UM1)-Université de Montpellier (UM), BUISINE, Soline, Integrative Biology of Emerging Infectious Diseases - - IBEID2010 - ANR-10-LABX-0062 - LABX - VALID, LES FACTEURS DE L'HÔTE ASSOCIÉS À LA NEUROPATHOGÉNICITÉ DU VIRUS ZIKA - - ZIKAHOST2017 - ANR-17-CE15-0029 - AAPG2017 - VALID, and Institut Convergences pour l'étude de l'Emergence des Pathologies au Travers des Individus et des populatiONs - - INCEPTION2016 - ANR-16-CONV-0005 - CONV - VALID

Subjects
Male, Permissiveness, viruses, Viral pathogenesis, Muscle Proteins, Host-Derived Cellular Factors, Alphavirus, Viral Nonstructural Proteins, Virus Replication, medicine.disease_cause, Virus, Myoblasts, Mice, 03 medical and health sciences, medicine, Animals, Humans, O'nyong-nyong Virus, Chikungunya, Cells, Cultured, 030304 developmental biology, Host factor, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, 0303 health sciences, Multidisciplinary, biology, 030306 microbiology, Intracellular Signaling Peptides and Proteins, virus diseases, Fibroblasts, LIM Domain Proteins, biology.organism_classification, Virology, FHL1, 3. Good health, HEK293 Cells, Viral replication, Host-Pathogen Interactions, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Chikungunya Fever, RNA, Viral, Female, Chikungunya virus, Protein Binding
Abstract

Chikungunya virus (CHIKV) is a re-emerging alphavirus that is transmitted to humans by mosquito bites and causes musculoskeletal and joint pain1,2. Despite intensive investigations, the human cellular factors that are critical for CHIKV infection remain unknown, hampering the understanding of viral pathogenesis and the development of anti-CHIKV therapies. Here we identified the four-and-a-half LIM domain protein 1 (FHL1)3 as a host factor that is required for CHIKV permissiveness and pathogenesis in humans and mice. Ablation of FHL1 expression results in the inhibition of infection by several CHIKV strains and o’nyong-nyong virus, but not by other alphaviruses and flaviviruses. Conversely, expression of FHL1 promotes CHIKV infection in cells that do not normally express it. FHL1 interacts directly with the hypervariable domain of the nsP3 protein of CHIKV and is essential for the replication of viral RNA. FHL1 is highly expressed in CHIKV-target cells and is particularly abundant in muscles3,4. Dermal fibroblasts and muscle cells derived from patients with Emery–Dreifuss muscular dystrophy that lack functional FHL15 are resistant to CHIKV infection. Furthermore, CHIKV infection is undetectable in Fhl1-knockout mice. Overall, this study shows that FHL1 is a key factor expressed by the host that enables CHIKV infection and identifies the interaction between nsP3 and FHL1 as a promising target for the development of anti-CHIKV therapies. FHL1 is a key factor expressed by humans and mice that is required for chikungunya virus infection and is therefore a promising target for the development of therapies against chikungunya virus.

Published
2019

21. Evaluation of a Trio Toscana Virus Real-Time RT-PCR Assay Targeting Three Genomic Regions within Nucleoprotein Gene

Author

Xavier de Lamballerie, Sara Sanbonmatsu-Gámez, Irene Pedrosa-Corral, Rémi N. Charrel, Alessandra Falchi, Mercedes Pérez-Ruiz, Laura Pezzi, Laurence Thirion, [Thirion,L, Pezzi,L, de Lamballerie,X, Charrel,RN] Unité des Virus Émergents (UVE: Aix-Marseille Univ-IRD 190-Inserm 1207-IHU Méditerranée Infection), Marseille, France. [Pezzi,L, Falchi,A] UR7310, Laboratoire de Virologie, Université de Corse-Inserm, Corte, France. [Pedrosa-Corral,I, Sanbonmatsu-Gamez,S, Perez-Ruiz,M] Servicio de Microbiología, Hospital Universitario Virgen de las Nieves, Instituto de Investigación Biosanitaria ibs.Granada, Granada, Spain., This study was partially funded by (i) the 'European Virus Archive Global' (EVA GLOBAL) project H2020-INFRAIA-2019 program, Project No 871029, and (ii) 'Emerging Viral Diseases—Expert Laboratory Network' (EVDLabNET).

Subjects
Microbiology (medical), Phlebovirus, Organisms::Viruses::RNA Viruses::Bunyaviridae::Phlebovirus::Sandfly fever Naples virus [Medical Subject Headings], Information Science::Information Science::Medical Informatics::Medical Informatics Applications::Information Storage and Retrieval::Data Mining [Medical Subject Headings], In silico, lcsh:Medicine, Phenuiviridae, Diseases::Bacterial Infections and Mycoses::Central Nervous System Infections [Medical Subject Headings], Biology, Mediterranean, Arbovirus, Article, Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::Humans [Medical Subject Headings], 03 medical and health sciences, Chemicals and Drugs::Nucleic Acids, Nucleotides, and Nucleosides::Nucleic Acids::RNA [Medical Subject Headings], medicine, diagnostics, Immunology and Allergy, Meningitis, Rna targeting, Diagnostics, Molecular Biology, Gene, 030304 developmental biology, 0303 health sciences, General Immunology and Microbiology, 030306 microbiology, Toscana virus, Diagnóstico, lcsh:R, Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Nucleoproteins [Medical Subject Headings], meningitis, Phlebotomus, medicine.disease, biology.organism_classification, Virology, 3. Good health, Nucleoprotein, Infectious Diseases, Real-time polymerase chain reaction, arbovirus, Sand fly, Psychodidae, sand fly
Abstract

Toscana virus (TOSV) can cause central nervous system infections in both residents of and travelers to Mediterranean countries. Data mining identified three real-time RT-qPCR assays for detecting TOSV RNA targeting non-overlapping regions in the nucleoprotein gene. Here, they were combined to create a multi-region assay named Trio TOSV RT-qPCR consisting of six primers and three probes. In this study, (i) we evaluated in silico the three RT-qPCR assays available in the literature for TOSV detection, (ii) we combined the three systems to create the Trio TOSV RT-qPCR, (iii) we assessed the specificity and sensitivity of the three monoplex assays versus the Trio TOSV RT-qPCR assay, and (iv) we compared the performance of the Trio TOSV RT-qPCR assay with one of the reference monoplex assays on clinical samples. In conclusion, the Trio TOSV RT-qPCR assay performs equally or better than the three monoplex assays, therefore, it provides a robust assay that can be used for both research and diagnostic purposes.

Published
2021

22. Zika Virus Seroprevalence in Two Districts of Chincha, Ica, Peru: A Cross-Sectional Study

Author

Eduardo Gotuzzo, Thomas Jaenisch, Rodrigo Cachay, Laura Pezzi, Luis Suarez-Ognio, Xiomara Merino, Michael Talledo, Alvaro Schwalb, Xavier de Lamballerie, J. Gonzalo Acevedo-Rodriguez, and Humberto Guerra

Subjects
Adult, Male, Multivariate analysis, Cross-sectional study, 030231 tropical medicine, Enzyme-Linked Immunosorbent Assay, Antibodies, Viral, Article, Herd immunity, Zika virus, Serology, Chincha, 03 medical and health sciences, symbols.namesake, Young Adult, 0302 clinical medicine, Neutralization Tests, Seroepidemiologic Studies, Virology, Peru, Seroprevalence, Humans, 030212 general & internal medicine, Poisson regression, Ica, Retrospective Studies, 2. Zero hunger, biology, Zika Virus Infection, Zika Virus Seroprevalence, Outbreak, Zika Virus, biology.organism_classification, 3. Good health, Cross-Sectional Study, Infectious Diseases, Geography, Cross-Sectional Studies, symbols, Parasitology, Female, Demography
Abstract

In 2017, a major outbreak of Zika virus (ZIKV) infection took place in Chincha Province, Peru, where arboviral circulation had never been reported before. We conducted a cross-sectional survey (March–May 2019) in two districts of Chincha Province: Pueblo Nuevo and Chincha Baja. We included residents who were 20 to 40 years old and who had lived in these districts for at least 1 year. Serological testing combined screening with a commercial NS1 protein-based Zika IgG ELISA, and confirmation by a cytopathic effect-based virus neutralization test (VNT). Prevalence ratios (PRs) were calculated using Poisson regression with robust error variance. Four hundred participants, divided equally among districts, were enrolled. Anti-ZIKV IgG ELISA was positive for 42 participants (10.5%) and borderline for 12 (3%). Fifty-two of these 54 samples were confirmed positive by ZIKV VNT (13% of the total population). The Pueblo Nuevo district exhibited a greater ZIKV seroprevalence based on VNT results than the Chincha Baja district (23.5% versus 2.5%), with participants from the Pueblo Nuevo district being 9.4 times more likely to have a positive ZIKV VNT result. Average monthly income greater than the minimum wage and adequate water storage were found to be protective factors (PR, 0.29 and 0.24, respectively). In multivariate analysis, living in the Pueblo Nuevo district and a personal history of fever and rash were strong predictors of ZIKV positivity by VNT. The low ZIKV seroprevalence should prompt health authorities to stimulate interventions to prevent potential future outbreaks. In the Pueblo Nuevo district, the seroprevalence was greater but presumably not sufficient to ensure protective herd immunity.

Published
2021
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23. Detection of Two Species of the Genus Parapoxvirus (Bovine Papular Stomatitis Virus and Pseudocowpox Virus) in Ticks Infesting Cattle in Burkina Faso

Author

Abel S. Biguezoton, Léa Luciani, Adrien Marie Gaston Belem, Laurence Thirion, Laura Pezzi, Rémi N. Charrel, Claude Saegerman, Achille S. Ouedraogo, Olivier M. Zannou, Laetitia Lempereur, Centre international de recherche-développement sur l'élevage en zone sub-humide (CIRDES), Faculty of Veterinary Medicine, Center for Fundamental and Applied Research for Animal and Health (FARAH), Laboratory of Parasitology and Parasitic Diseases, Université de Liège, Unité des Virus Emergents (UVE), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Research Unit in Epidemiology and Risk Analysis Applied to the Veterinary Sciences (UREAR-ULg), Department of Infectious and Parasitic Diseases, Center for Fundamental and Applied Research for Animal and Health (FARAH), Faculty of Veterinary Medicine, EA Bioscope Corse Méditerranée : Dynamique des infections virales en milieu insulaire, Université Pascal Paoli (UPP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut du Développement Rural, Université Polytechnique Nazi Boni Bobo-Dioulasso (UNB), BUISINE, Soline, and Aix Marseille Université (AMU)-Institut de Recherche pour le Développement (IRD)-Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
0301 basic medicine, Microbiology (medical), Veterinary medicine, 040301 veterinary sciences, viruses, [SDV]Life Sciences [q-bio], Population, virus, Tick, Pseudocowpox, cattle ticks, Microbiology, Article, 0403 veterinary science, 03 medical and health sciences, Bovine papular stomatitis, Virology, parasitic diseases, West Africa, Burkina Faso, medicine, education, lcsh:QH301-705.5, 2. Zero hunger, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, education.field_of_study, pseudocowpox, biology, 04 agricultural and veterinary sciences, biology.organism_classification, medicine.disease, 3. Good health, [SDV] Life Sciences [q-bio], Pseudocowpox Virus, 030104 developmental biology, arbovirus, Phlebovirus, lcsh:Biology (General), poxviridae, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, bovine papular stomatitis, Hyalomma, Amblyomma variegatum
Abstract

International audience; The molecular identification of arboviruses in West Africa is of particular interest, due to their zoonotic potential in a population living in close contact with livestock, and in a region where the livestock migration across borders raises the risk of diseases infection and dissemination. The aim of the study was the screening of potential circulating arboviruses and the assessment of their zoonotic implications. Therefore, ticks were collected on cattle located in three provinces of eastern Burkina Faso. Tick pools were tested using a panel of genus-specific real-time assays targeting conserved regions of parapoxvirus, orthopoxvirus, flavivirus and phlebovirus. On the 26 farms visited, a total of 663 ticks were collected. Four genera and six tick species were morphologically identified, with Amblyomma variegatum and Hyalomma spp. being the most represented species. No arboviruses were found. However, this study highlights the presence of pseudocowpox virus (8.2%) and bovine papular stomatitis virus (5.8%) among the positive tick pools. BPSV positive ticks were found in herds sharing water and pastures resources and with a history of seasonal transhumance. Therefore, common grazing and the seasonal transhumance are likely to support the transmission of the virus. This could have important health and economic impacts, especially regarding transboundary cattle movements

Published
2020

24. Lyophilized Matrix Containing Ready-to-Use Primers and Probe Solution for Standardization of Real-Time PCR and RT-qPCR Diagnostics in Virology

Author

Mhammed Touinssi, Xavier de Lamballerie, Laura Pezzi, Audrey Dubot-Pérès, Laurence Thirion, Iban Corcostegui, Rémi N. Charrel, Unité des Virus Emergents (UVE), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Hospitalier Universitaire Méditerranée Infection (IHU Marseille), Laboratoire de Virologie [UNIV Corse-Inserm] (EA7310), Université Pascal Paoli (UPP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Établissement Français du Sang Alpes-Méditerranée (EFS Alpes-Méditerranée), University of Florida [Gainesville] (UF), and European Project: 653316,H2020,H2020-INFRAIA-2014-2015,EVAg(2015)

Subjects
0301 basic medicine, Rift Valley fever phlebovirus, diagnosis, [SDV]Life Sciences [q-bio], 030106 microbiology, TaqMan, lcsh:QR1-502, lyophilization, virus, Real-Time Polymerase Chain Reaction, Sensitivity and Specificity, Article, lcsh:Microbiology, epidemic, Single tube, Matrix (chemical analysis), 03 medical and health sciences, Freeze-drying, Virology, ComputingMilieux_MISCELLANEOUS, DNA Primers, Chromatography, Reverse Transcriptase Polymerase Chain Reaction, Chemistry, emerging, 3. Good health, Oligonucleotide primers, Freeze Drying, 030104 developmental biology, Infectious Diseases, Real-time polymerase chain reaction, PCR, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Molecular Diagnostic Techniques, Viruses, freeze-drying, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Ready to use, DNA Probes, pathogen
Abstract

Real-time molecular techniques have become the reference methods for direct diagnosis of pathogens. The reduction of steps is a key factor in order to decrease the risk of human errors resulting in invalid series and delayed results. We describe here a process of preparation of oligonucleotide primers and hydrolysis probe in a single tube at predefined optimized concentrations that are stabilized via lyophilization (Lyoph-P&amp, P). Lyoph-P&amp, P was compared versus the classic protocol using extemporaneously prepared liquid reagents using (i) sensitivity study, (ii) long-term stability at 4 &deg, C, and (iii) long-term stability at 37 &deg, C mimicking transportation without cold chain. Two previously published molecular assays were selected for this study. They target two emerging viruses that are listed on the blueprint of the WHO as to be considered for preparedness and response actions: chikungunya virus (CHIKV) and Rift Valley fever phlebovirus (RVFV). Results of our study demonstrate that (i) Lyoph-P&amp, P is stable for at least 4 days at 37 &deg, C supporting shipping without the need of cold chain, (ii) Lyoph-P&amp, P rehydrated solution is stable at +4 &deg, C for at least two weeks, (iii) sensitivity observed with Lyoph-P&amp, P is at least equal to, often better than, that observed with liquid formulation, (iv) validation of results observed with low-copy specimens is rendered easier by higher fluorescence level. In conclusion, Lyoph-P&amp, P holds several advantages over extemporaneously preparer liquid formulation that merit to be considered when a novel real-time molecular assay is implemented in a laboratory in charge of routine diagnostic activity.

Published
2020

25. Brainstem Correlates of a Cold Pressor Test Measured by Ultra-High Field fMRI

Author

Domenica Bueti, Mariëlle C. Hendriks-Balk, Fatma Megdiche, Dimitri Van De Ville, Olivier Reynaud, Laura Pezzi, Sandra Da Costa, and Grégoire Wuerzner

Subjects
Brain activity and meditation, blood pressure, BOLD, brainstem, cold pressor test, fMRI, sympathetic nervous system, 030204 cardiovascular system & hematology, 0302 clinical medicine, blood-pressure, sites, Original Research, CIBM-AIT, medicine.diagnostic_test, General Neuroscience, sympathetic-nerve activity, Cold pressor test, Neurogenic hypertension, medicine.anatomical_structure, supplementary, FMRI, Cardiology, Blood pressure, Brainstem, Motor cortex, medicine.medical_specialty, hypertension, drive, challenges, Statistical parametric mapping, Insular cortex, ddc:616.0757, lcsh:RC321-571, 03 medical and health sciences, Internal medicine, medicine, lcsh:Neurosciences. Biological psychiatry. Neuropsychiatry, business.industry, Settore M-PSI/02 - Psicobiologia e Psicologia Fisiologica, Sympathetic nervous system, responses, activation, Functional magnetic resonance imaging, business, 030217 neurology & neurosurgery, Neuroscience
Abstract

Modern imaging techniques such as blood oxygen level-dependent (BOLD) functional magnetic resonance imaging (fMRI) allow the non-invasive and indirect measurement of brain activity. Whether changes in signal intensity can be detected in small brainstem regions during a cold pressor test (CPT) has not been explored thoroughly. The aim of this study was to measure jointly whole brain and brainstem BOLD signal intensity changes in response to a modified CPT. Methods: BOLD fMRI was measured in healthy normotensive participants during unblinded a randomized crossover study (modified CPT vs. control test) using ultra-high field 7 Tesla MRI scanner. Data were analyzed using Statistical Parametric Mapping (SPM) in a whole-brain approach, and with a brainstem-specific analysis using the spatially unbiased infra-tentorial template (SUIT) toolbox. Blood pressure (BP) and hormonal responses (norepinephrine and epinephrine levels) were also measured. Paired t-test statistics were used to compare conditions. Results: Eleven healthy volunteersparticipants (6 womanwomen, mean age 28 ± 8.9 years) were enrolledanalyzed after giving written, informed consent. Mean arterial BP increased from 834 ± 121 mm Hg to 876 ± 12 mm Hg (p=0.00974) during the CPT. Whole-brain analysis revealed significant activations linked to the CPT in the right supplementary motor cortex, midcingulate (bilateral) and the right anterior insular cortex. The brainstem-specific analysis showed significant activations in the dorsal medulla. Conclusions: Changes in BOLD fMRI signal intensity in brainstem regions during a CPT can be detected, and show an increased response during a cold stress in healthy volunteers. Consequentlyvolunteers. Consequently, BOLD fMRI at 7T is a promising tool to explore and acquire new insights in the comprehension of neurogenic hypertension.

Published
2020

26. FHL1 is a key player of chikungunya virus tropism and pathogenesis

Author

Beate M. Kümmerer, Laura Pezzi, Mohamed Lamine Hafirassou, Lucie Gueneau, Constance Delaugerre, Gisèle Bonne, Thibaud Goupil, Vasiliya Kril, Monsef Benkirane, Julien Burlaud-Gaillard, Lucie Bonnet-Madin, R. Juntas-Morales, Anne Bertrand Legout, Thérèse Couderc, Laurent Meertens, Etienne Simon-Loriere, Philippe Roingeard, Ali Amara, Marc Lecuit, Rabah Ben Yaou, Sophia Rafasse, Alexis Brugier, Xavier de Lamballerie, Pierre-Olivier Vidalain, Cécile Doyen, Athena Labeau, Génomes, biologie cellulaire et thérapeutiques (GenCellDi (U944 / UMR7212)), Collège de France (CdF (institution))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Biologie des Infections - Biology of Infection, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Universität Bonn = University of Bonn, Génomique évolutive des virus à ARN - Evolutionary genomics of RNA viruses, Institut Pasteur [Paris] (IP), Morphogénèse et antigénicité du VIH et du virus des Hépatites (MAVIVH - U1259 Inserm - CHRU Tours ), Centre Hospitalier Régional Universitaire de Tours (CHRU Tours)-Université de Tours (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de génétique humaine (IGH), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Unité des Virus Emergents (UVE), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Modélisation et Immunologie pour la Thérapie (CBMIT), Université Paris Descartes - Paris 5 (UPD5), Centre de recherche en Myologie – U974 SU-INSERM, Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier), Hopital Saint-Louis [AP-HP] (AP-HP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Université Paris Cité (UPCité), CHU Necker - Enfants Malades [AP-HP], Service des Maladies infectieuses et tropicales [CHU Necker], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Imagine - Institut des maladies génétiques (IHU) (Imagine - U1163), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Génomes, biologie cellulaire et thérapeutiques (GenCellDi (UMR_S_944)), Collège de France (CdF (institution))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), University of Bonn, Institut Pasteur [Paris], Aix Marseille Université (AMU)-Institut de Recherche pour le Développement (IRD)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Paris (UP), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Paris (UP), DIAKITE, andrée, Université de Tours-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Hospitalier Régional Universitaire de Tours (CHRU TOURS), and Centre de Recherche en Myologie

Subjects
0106 biological sciences, 0301 basic medicine, viruses, [SDV]Life Sciences [q-bio], Muscle Proteins, Pathogenesis, MESH: Chikungunya Fever, Viral Nonstructural Proteins, Virus Replication, medicine.disease_cause, 01 natural sciences, Mice, Four and a Half LIM domains 1 Tropism, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, MESH: Animals, Chikungunya, Host factor, [SDV.MHEP.ME] Life Sciences [q-bio]/Human health and pathology/Emerging diseases, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, [SDV.MHEP.ME]Life Sciences [q-bio]/Human health and pathology/Emerging diseases, [SDV.MHEP] Life Sciences [q-bio]/Human health and pathology, biology, Intracellular Signaling Peptides and Proteins, virus diseases, General Medicine, MESH: Tropism, LIM Domain Proteins, 3. Good health, [SDV] Life Sciences [q-bio], [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Chikungunya virus, MESH: LIM Domain Proteins, Viral protein, Virus infection, Alphavirus, Tropism, Virus, 03 medical and health sciences, MESH: Muscle Proteins, MESH: Intracellular Signaling Peptides and Proteins, medicine, Animals, [SDV.MP] Life Sciences [q-bio]/Microbiology and Parasitology, MESH: Mice, Aedes, MESH: Virus Replication, MESH: Chikungunya virus, biology.organism_classification, Virology, 030104 developmental biology, Infectious disease (medical specialty), Chikungunya Fever, MESH: Viral Nonstructural Proteins, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology, 010606 plant biology & botany
Abstract

Chikungunya is an infectious disease caused by the chikungunya virus (CHIKV), an alphavirus transmitted to humans by Aedes mosquitoes, and for which there is no licensed vaccine nor antiviral treatments. By using a loss-of-function genetic screen, we have recently identified the FHL1 protein as an essential host factor for CHIKV tropism and pathogenesis. FHL1 is highly expressed in muscles cells and fibroblasts, the main CHIKV-target cells. FHL1 interacts with the viral protein nsP3 and plays a critical role in CHIKV genome amplification. Experiments in vivo performed in FHL1-deficient mice have shown that these animals are resistant to infection and do not develop muscular lesions. Altogether these observations, published in the journal Nature [1], show that FHL1 is a key host factor for CHIKV pathogenesis and identify the interaction between FHL1 and nsP3 as a promising target for the development of new antiviral strategies., Le chikungunya est une maladie infectieuse causée par le virus chikungunya (CHIKV), un alphavirus transmis à l’Homme par les moustiques Aedes et contre lequel il n’existe ni vaccin, ni traitements antiviraux. En utilisant une approche de crible génétique par perte de fonction, nous avons récemment identifié la protéine FHL1 comme un facteur cellulaire essentiel pour le tropisme et la pathogénèse du CHIKV. FHL1 est une molécule présente majoritairement dans les cellules musculaires et les fibroblastes, les cibles privilégiées de CHIKV. FHL1 interagit avec la protéine virale nsP3 et joue un rôle décisif dans le mécanisme d’amplification du génome de CHIKV. Des expériences in vivo chez des souris déficientes pour FHL1 ont montré que ces animaux sont résistants à l’infection et ne développent pas de lésions musculaires. L’ensemble de ces observations publiées dans la revue Nature [1] montrent que FHL1 est un facteur cellulaire clé pour la pathogénèse de CHIKV et identifient l’interaction entre FHL1 et nsp3 comme une cible prometteuse pour le développement de nouvelles stratégies antivirales

Published
2020

27. GloPID-R report on chikungunya, o'nyong-nyong and Mayaro virus, part 5: Entomological aspects

Author

Thomas Jaenisch, Anubis Vega-Rúa, Patrícia Brasil, M. G. Rosa-Freitas, Michael A. Drebot, P. Gallian, Sébastien Boyer, Maria Rios, Amadou A. Sall, Laura Pezzi, Anna-Bella Failloux, Angelle Desiree LaBeaud, Lisa F. P. Ng, Michael S. Diamond, Ricardo Lourenço-de-Oliveira, Guilherme S. Ribeiro, Scott C. Weaver, Mariama Dalanda Diallo, Nikos Vasilakis, A. J. Rodriguez-Morales, Alain Kohl, François Simon, Michael P. Busch, André Siqueira, Chantal Reusken, Jan Felix Drexler, Johan Neyts, Graham Simmons, X. de Lamballerie, Marc Lecuit, Unité des Virus Emergents (UVE), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM), EA Bioscope Corse Méditerranée : Dynamique des infections virales en milieu insulaire, Université Pascal Paoli (UPP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Pasteur de Dakar, Réseau International des Instituts Pasteur (RIIP), Fundação Oswaldo Cruz (FIOCRUZ), Institut Pasteur de la Guadeloupe, Agency for science, technology and research [Singapore] (A*STAR), Institut Pasteur du Cambodge, Charité - UniversitätsMedizin = Charité - University Hospital [Berlin], German Centre for Infection Research (DZIF), The University of Texas Medical Branch (UTMB), MRC - University of Glasgow Centre for Virus Research, Arbovirus et Insectes Vecteurs - Arboviruses and Insect Vectors, Institut Pasteur [Paris] (IP), University of California [San Francisco] (UC San Francisco), University of California (UC), Washington University School of Medicine in St. Louis, Washington University in Saint Louis (WUSTL), Public Health Agency of Canada, Etablissement Français du Sang - Alpes-Méditerranée (EFS - Alpes-Méditerranée), Etablissement Français du Sang, Heidelberg University Hospital [Heidelberg], Stanford School of Medicine [Stanford], Stanford Medicine, Stanford University-Stanford University, Biologie des Infections - Biology of Infection, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris Descartes - Paris 5 (UPD5), Service des Maladies infectieuses et tropicales [CHU Necker], CHU Necker - Enfants Malades [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Catholic University of Leuven - Katholieke Universiteit Leuven (KU Leuven), Rega Institute for Medical Research [Leuven, België], National Institute for Public Health and the Environment [Bilthoven] (RIVM), Erasmus University Medical Center [Rotterdam] (Erasmus MC), Universidade Federal da Bahia (UFBA), Instituto Gonçalo Moniz / Gonçalo Moniz Research Centre - Fiocruz Bahia [Salvador, Brésil] (IGM), Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), U.S. Food and Drug Administration (FDA), Universidad Tecnológica de Pereira [Colombie] (UTP), Hôpital d'Instruction des Armées Laveran, Service de Santé des Armées, BUISINE, Soline, Virology, Fundação Oswaldo Cruz / Oswaldo Cruz Foundation (FIOCRUZ), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pascal Paoli (UPP), Institut Pasteur [Paris], University of California [San Francisco] (UCSF), University of California, and Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
0301 basic medicine, Primates, Research Report, GloPID-R chikungunya, o'nyong-nyong and Mayaro virus Working Group, Aedes albopictus, [SDV]Life Sciences [q-bio], 030106 microbiology, Aedes aegypti, Mosquito Vectors, medicine.disease_cause, 03 medical and health sciences, Aedes, Virology, Anopheles, medicine, Animals, Humans, O'nyong-nyong Virus, Chikungunya, Haemagogus, [SDV.MP] Life Sciences [q-bio]/Microbiology and Parasitology, Pharmacology, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, biology, Alphavirus Infections, Yellow fever, Outbreak, virus diseases, Central America, biology.organism_classification, medicine.disease, 3. Good health, [SDV] Life Sciences [q-bio], 030104 developmental biology, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Infectious disease (medical specialty), Africa, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Enzootic, Chikungunya Fever, Chikungunya virus
Abstract

The GloPID-R (Global Research Collaboration for Infectious Disease Preparedness) chikungunya (CHIKV), o'nyong-nyong (ONNV) and Mayaro virus (MAYV) Working Group has been established to investigate natural history, epidemiology and clinical aspects of infection by these viruses. Here, we present a report dedicated to entomological aspects of CHIKV, ONNV and MAYV. Recent global expansion of chikungunya virus has been possible because CHIKV established a transmission cycle in urban settings using anthropophilic vectors such as Aedes albopictus and Aedes aegypti. MAYV and ONNV have a more limited geographic distribution, being confined to Africa (ONNV) and central-southern America (MAYV). ONNV is probably maintained through an enzootic cycle that has not been characterized yet, with Anopheles species as main vectors and humans as amplification hosts during epidemics. MAYV is transmitted by Haemagogus species in an enzootic cycle using non-human primates as the main amplification and maintenance hosts, and humans becoming sporadically infected when venturing in or nearby forest habitats. Here, we focused on the transmission cycle and natural vectors that sustain circulation of these viruses in their respective locations. The knowledge of the natural ecology of transmission and the capacity of different vectors to transmit these viruses is crucial to understand CHIKV emergence, and to assess the risk that MAYV and ONNV will expand on wide scale using anthropophilic mosquito species not normally considered primary vectors. Finally, the experts identified knowledge gaps and provided adapted recommendations, in order to address future entomological investigations in the right direction. ispartof: ANTIVIRAL RESEARCH vol:174 ispartof: location:Netherlands status: published

Published
2019

28. GloPID-R report on chikungunya, o'nyong-nyong and Mayaro virus, part 2: Epidemiological distribution of o'nyong-nyong virus

Author

M. G. Rosa-Freitas, Sébastien Boyer, A. J. Rodriguez-Morales, Jan Felix Drexler, Thomas Jaenisch, Patrícia Brasil, Johan Neyts, François Simon, Anna-Bella Failloux, de Lamballerie X, Lisa F. P. Ng, Michael A. Drebot, Pierre Gallian, Guilherme S. Ribeiro, Maria Rios, Amadou A. Sall, Chantal Reusken, Marc Lecuit, Scott C. Weaver, Michael P. Busch, Nikos Vasilakis, André Siqueira, A. Vega Rua, Angelle Desiree LaBeaud, Mawlouth Diallo, Michael S. Diamond, Alain Kohl, Ricardo Lourenço-de-Oliveira, Laura Pezzi, Graham Simmons, Unité des Virus Emergents (UVE), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de Virologie [UNIV Corse-Inserm] (EA7310), Université Pascal Paoli (UPP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Stanford School of Medicine [Stanford], Stanford Medicine, Stanford University-Stanford University, National Institute for Public Health and the Environment [Bilthoven] (RIVM), Erasmus University Medical Center [Rotterdam] (Erasmus MC), Charité - UniversitätsMedizin = Charité - University Hospital [Berlin], Humboldt University Of Berlin, Berlin Institute of Health (BIH), The University of Texas Medical Branch (UTMB), Institut Pasteur de Dakar, Réseau International des Instituts Pasteur (RIIP), Hôpital d'Instruction des Armées Laveran, Service de Santé des Armées, Heidelberg University Hospital [Heidelberg], Etablissement Français du Sang - Alpes-Méditerranée (EFS - Alpes-Méditerranée), Etablissement Français du Sang, Arbovirus et Insectes Vecteurs - Arboviruses and Insect Vectors, Institut Pasteur [Paris] (IP), Institut Pasteur du Cambodge, Fundação Oswaldo Cruz / Oswaldo Cruz Foundation (FIOCRUZ), University of California [San Francisco] (UC San Francisco), University of California (UC), Washington University in Saint Louis (WUSTL), Public Health Agency of Canada, MRC - University of Glasgow Centre for Virus Research, Université Paris Descartes - Paris 5 (UPD5), CHU Necker - Enfants Malades [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Biologie des Infections - Biology of Infection, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Imagine - Institut des maladies génétiques (IHU) (Imagine - U1163), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Catholic University of Leuven - Katholieke Universiteit Leuven (KU Leuven), Agency for science, technology and research [Singapore] (A*STAR), Universidade Federal da Bahia (UFBA), U.S. Food and Drug Administration (FDA), Universidad Tecnológica de Pereira [Colombie] (UTP), Institut Pasteur de la Guadeloupe, Publication fees were covered by the Global Research Collaboration for Infectious Disease Preparedness (GloPID-R)., Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pascal Paoli (UPP), Humboldt-Universität zu Berlin, Institut Pasteur [Paris], Fundação Oswaldo Cruz (FIOCRUZ), University of California [San Francisco] (UCSF), University of California, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Paris (UP), Virology, and DEMESLAY GOUGAM, MARIE

Subjects
0301 basic medicine, GloPID-R chikungunya, o'nyong-nyong and Mayaro virus Working Group, Genes, Viral, Iron, 030106 microbiology, Prevalence, Alphavirus, medicine.disease_cause, Disease Outbreaks, 03 medical and health sciences, Seroepidemiologic Studies, Virology, Environmental health, medicine, Seroprevalence, Animals, Humans, O'nyong-nyong Virus, Serologic Tests, Chikungunya, Phylogeny, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Pharmacology, Molecular epidemiology, Alphavirus Infections, Outbreak, 3. Good health, 030104 developmental biology, Geography, Infectious disease (medical specialty), [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Africa, Chikungunya Fever, Chikungunya virus
Abstract

The GloPID-R (Global Research Collaboration for Infectious Disease Preparedness) chikungunya (CHIKV), o'nyong-nyong (ONNV) and Mayaro virus (MAYV) Working Group has been established to identify gaps of knowledge about the natural history, epidemiology and medical management of infection by these viruses, and to provide adapted recommendations for future investigations. Here, we present a report dedicated to ONNV epidemiological distribution. Two large-scale ONNV outbreaks have been identified in Africa in the last 60 years, interspersed with sporadic serosurveys and case reports of returning travelers. The assessment of the real scale of ONNV circulation in Africa remains a difficult task and surveillance studies are necessary to fill this gap. The identification of ONNV etiology is made complicated by the absence of multiplex tools in co-circulation areas and that of reference standards, as well as the high cross-reactivity with related pathogens observed in serological tests, in particular with CHIKV. This is a specific obstacle for seroprevalence studies, that necessitate an improvement of serological tools to provide robust results. The scarcity of existent genetic data currently limits molecular epidemiology studies. ONNV epidemiology would also benefit from reinforced entomological and environmental surveillance. Finally, the natural history of the disease deserves to be further investigated, with a specific attention paid to long-term complications. Considering our incomplete knowledge on ONNV distribution, GloPID-R CHIKV, ONNV and MAYV experts recommend that a major effort should be done to fill existing gaps. ispartof: ANTIVIRAL RESEARCH vol:172 ispartof: location:Netherlands status: published

Published
2019

29. Development and Evaluation of a Duo Chikungunya Virus Real-Time RT-PCR Assay Targeting Two Regions within the Genome

Author

Rémi N. Charrel, Laurence Thirion, Iban Corcostegui, Xavier de Lamballerie, Alessandra Falchi, Laura Pezzi, Audrey Dubot-Pérès, Unité des Virus Emergents (UVE), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de Virologie [UNIV Corse-Inserm] (EA7310), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pascal Paoli (UPP), Emerging Pathogens Institute [Gainesville, FL, USA], University of Florida [Gainesville] (UF), This research was partly funded by (i) the European Virus Archive (European Union Seventh Framework Programme under grant agreement no. 228292) (http://www.european-virus-archive.com/), (ii) the European Virus Archive goes Global (EVAg) project that has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 653316, (iii) Université de Corse Pasquale Paoli, Corte, France (UCPP), (iv) Emerging Viral Diseases-Expert Laboratory Network (EVD-LabNet). L.P. is a PhD student granted by UCPP and INSERM., European Project: 228292,EC:FP7:INFRA,FP7-INFRASTRUCTURES-2008-1,EVA(2009), European Project: 653316,H2020,H2020-INFRAIA-2014-2015,EVAg(2015), NOUGAIREDE, Antoine, European Virus Archive - EVA - - EC:FP7:INFRA2009-01-01 - 2014-12-31 - 228292 - VALID, and European Virus Archive goes global - EVAg - - H20202015-04-01 - 2019-03-31 - 653316 - VALID

Subjects
0301 basic medicine, In silico, 030106 microbiology, lcsh:QR1-502, Alphavirus, Genome, Viral, Biology, medicine.disease_cause, Genome, Arbovirus, Virus, lcsh:Microbiology, Article, epidemic, Evolution, Molecular, 03 medical and health sciences, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Virology, medicine, Togaviridae, emergence, Humans, Chikungunya, arthropod-borne, Genetics, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, [SDV.MHEP.ME] Life Sciences [q-bio]/Human health and pathology/Emerging diseases, fever, [SDV.MHEP.ME]Life Sciences [q-bio]/Human health and pathology/Emerging diseases, outbreak, Reverse Transcriptase Polymerase Chain Reaction, Point mutation, virus diseases, medicine.disease, biology.organism_classification, tropical disease, 030104 developmental biology, Infectious Diseases, arbovirus, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Mutation, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Chikungunya Fever, Chikungunya virus
Abstract

Laurence Thirion and Laura Pezzi contributed equally.; International audience; Chikungunya virus (CHIKV) re-emerged as a globalized health threat fifteen years ago. There are dozens of RT-PCR assays published. An inventory of the latter was made, and after in silico analysis, two assays were selected for their ability to detect strains belonging to the five CHIKV genetic lineages. They were combined in order to provide a robust assay not affected by genetic point mutations and the resulting Duo CHIKV real-time RT-PCR assay was compared to the two parental single-plex tests against five strains belonging to the five genetic lineages. The Duo CHIKV assay performed equally, or better, in terms of sensitivity, specificity, linearity and signal intensity. Dual-target assays are better suited for viruses having the propensity to evolve into new variants via point mutations or major sequence deletions/insertions. Here, we demonstrated that combining two single systems into a dual-target assay did not impair sensitivity and specificity, and proved a potent diagnostic tool to face a potential emergence of CHIKV variants by newly evolving mutations.

Published
2019

30. GloPID-R report on Chikungunya, O'nyong-nyong and Mayaro virus, part I: Biological diagnostics

Author

A. J. Rodriguez-Morales, M. G. Rosa-Freitas, Guilherme S. Ribeiro, Nikos Vasilakis, Lisa F. P. Ng, Mariama Dalanda Diallo, Scott C. Weaver, Patrícia Brasil, Jan Felix Drexler, Johan Neyts, Anna-Bella Failloux, François Simon, Alain Kohl, A. Vega Rua, Maria Rios, Michael P. Busch, Michael S. Diamond, André Siqueira, Michael A. Drebot, P. Gallian, Sébastien Boyer, Ricardo Lourenço-de-Oliveira, Chantal Reusken, Angelle Desiree LaBeaud, Thomas Jaenisch, Marc Lecuit, Laura Pezzi, X. de Lamballerie, Graham Simmons, Amadou A. Sall, Unité des Virus Emergents (UVE), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de Virologie [UNIV Corse-Inserm] (EA7310), Université Pascal Paoli (UPP)-Institut National de la Santé et de la Recherche Médicale (INSERM), National Institute for Public Health and the Environment [Bilthoven] (RIVM), Erasmus University Medical Center [Rotterdam] (Erasmus MC), The University of Texas Medical Branch (UTMB), Charité - UniversitätsMedizin = Charité - University Hospital [Berlin], Humboldt University Of Berlin, Berlin Institute of Health (BIH), German Centre for Infection Research (DZIF), University of California [San Francisco] (UC San Francisco), University of California (UC), Stanford School of Medicine [Stanford], Stanford Medicine, Stanford University-Stanford University, Washington University in Saint Louis (WUSTL), Public Health Agency of Canada, Fundação Oswaldo Cruz / Oswaldo Cruz Foundation (FIOCRUZ), Réseau International des Instituts Pasteur (RIIP), Universidade Federal da Bahia (UFBA), University of Glasgow, Biologie des Infections - Biology of Infection, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Imagine - Institut des maladies génétiques (IMAGINE - U1163), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), CHU Necker - Enfants Malades [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Université Paris Descartes - Paris 5 (UPD5), Agency for science, technology and research [Singapore] (A*STAR), Etablissement Français du Sang - Alpes-Méditerranée (EFS - Alpes-Méditerranée), Etablissement Français du Sang, Institut Pasteur du Cambodge, Institut Pasteur de Dakar, Arbovirus et Insectes Vecteurs - Arboviruses and Insect Vectors, Institut Pasteur [Paris] (IP), Heidelberg University Hospital [Heidelberg], Rega Institute for Medical Research [Leuven, België], Catholic University of Leuven - Katholieke Universiteit Leuven (KU Leuven), U.S. Food and Drug Administration (FDA), Universidad Tecnológica de Pereira [Colombie] (UTP), Hôpital d'Instruction des Armées Laveran, Service de Santé des Armées, Institut Pasteur de la Guadeloupe, Publication fees were covered by the Global Research Collaboration for Infectious Disease Preparedness (GloPID-R)., Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pascal Paoli (UPP), Humboldt-Universität zu Berlin, University of California [San Francisco] (UCSF), University of California, Fundação Oswaldo Cruz (FIOCRUZ), Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Pasteur [Paris], Virology, Charité - Universitätsmedizin Berlin / Charite - University Medicine Berlin, Humboldt Universität zu Berlin, Stanford University School of Medicine [Stanford], Stanford University [Stanford], Washington University in St Louis, Federal University of Bahia, Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Arbovirus et Insectes Vecteurs, and Rega Institute for Medical Research

Subjects
0301 basic medicine, medicine.medical_specialty, Genes, Viral, 030106 microbiology, Disease, Alphavirus, Mosquito Vectors, Cross Reactions, medicine.disease_cause, Antibodies, Viral, Communicable Diseases, Emerging, Virus, 03 medical and health sciences, Seroepidemiologic Studies, Virology, O'nyong-nyong, Epidemiology, medicine, Animals, Humans, O'nyong-nyong Virus, Chikungunya, Pathology, Molecular, Diagnostics, Phylogeny, Pharmacology, biology, business.industry, Alphavirus Infections, Outbreak, biology.organism_classification, Mayaro virus, 3. Good health, 030104 developmental biology, Cryoglobulinemia, Infectious disease (medical specialty), Preparedness, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, Chikungunya Fever, business, Chikungunya virus
Abstract

The GloPID-R (Global Research Collaboration for Infectious Disease Preparedness) Chikungunya (CHIKV), O'nyong-nyong (ONNV) and Mayaro virus (MAYV) Working Group is investigating the natural history, epidemiology and medical management of infection by these viruses, to identify knowledge gaps and to propose recommendations for direct future investigations and rectification measures. Here, we present the first report dedicated to diagnostic aspects of CHIKV, ONNV and MAYV. Regarding diagnosis of the disease at the acute phase, molecular assays previously described for the three viruses require further evaluation, standardized protocols and the availability of international standards representing the genetic diversity of the viruses. Detection of specific IgM would benefit from further investigations to clarify the extent of cross-reactivity among the three viruses, the sensitivity of the assays, and the possible interfering role of cryoglobulinaemia. Implementation of reference panels and external quality assessments for both molecular and serological assays is necessary. Regarding sero-epidemiological studies, there is no reported high-throughput assay that can distinguish among these different viruses in areas of potential co-circulation. New specific tools and/or improved standardized protocols are needed to enable large-scale epidemiological studies of public health relevance to be performed. Considering the high risk of future CHIKV, MAYV and ONNV outbreaks, the Working Group recommends that a major investigation should be initiated to fill the existing diagnostic gaps. ispartof: ANTIVIRAL RESEARCH vol:166 pages:66-81 ispartof: location:Netherlands status: published

Published
2019

31. Acquisition of respiratory and gastrointestinal pathogens among health care workers during the 2015 Hajj season

Author

Philippe Gautret, Hail M. Al-Abdely, Laura Pezzi, Abdulaziz Mushi, Rémi N. Charrel, Yara Yassin, Samir Benkouiten, Fuad Maashi, Badriah Alotaibi, Hanan H. Balkhy, Saber Yezli, Didier Raoult, Ministry of Health [Riyadh], King Saud Bin Abdulaziz University for Health Sciences [Riyadh] (KSAU-HS), Unité des Virus Emergents (UVE), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Vecteurs - Infections tropicales et méditerranéennes (VITROME), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut de Recherche Biomédicale des Armées [Brétigny-sur-Orge] (IRBA), Microbes évolution phylogénie et infections (MEPHI), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), COMBE, Isabelle, Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut de Recherche Biomédicale des Armées (IRBA), and King Saud Bin Abdulaziz University for Health Sciences

Subjects
Male, Epidemiology, health care facilities, manpower, and services, Cohort Studies, 0302 clinical medicine, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Health care, 030212 general & internal medicine, Respiratory system, Respiratory Tract Infections, Aged, 80 and over, [SDV.MHEP.ME] Life Sciences [q-bio]/Human health and pathology/Emerging diseases, 0303 health sciences, [SDV.MHEP.ME]Life Sciences [q-bio]/Human health and pathology/Emerging diseases, Transmission (medicine), Health Policy, Middle Aged, Gastroenteritis, 3. Good health, Religion, Infectious Diseases, Carrier State, Cohort, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Respiratory, Respiratory virus, Female, Adult, medicine.medical_specialty, Gastrointestinal, Infectious Disease Transmission, Patient-to-Professional, Health Personnel, education, Saudi Arabia, Gastrointestinal pathogens, Article, Young Adult, 03 medical and health sciences, Occupational Exposure, medicine, Humans, Aged, Carriage, 030306 microbiology, business.industry, Public Health, Environmental and Occupational Health, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, Crowding, Emergency medicine, Hajj, [SDV.MP.BAC] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, business, Multiplex Polymerase Chain Reaction
Abstract

International audience; Background: Data on the risk of transmission of infection to health care workers (HCWs) serving ill pilgrims during the Hajj is scarce.Methods: Two cohorts of HCWs, the first serving Hajj pilgrims in Mecca and the second serving patients in Al-Ahsa, were investigated for respiratory and gastrointestinal symptoms and pathogen carriage using multi-plex polymerase chain reaction before and after the 2015 Hajj.Results: A total of 211 HCWs were enrolled of whom 92 were exposed to pilgrims (Mecca cohort), whereas 119 were not exposed (Al-Ahsa cohort). Symptoms were observed only in HCWs from the Mecca cohort, with 29.3% experiencing respiratory symptoms during the Hajj period or in the subsequent days and 3.3% having gastrointestinal symptoms. Acquisition rates of at least 1 respiratory virus were 14.7% in the Mecca cohort and 3.4% in the Al-Ahsa cohort (P = .003). Acquisition rates of at least 1 respiratory bacterium were 11.8% and 18.6% in the Mecca and Al-Ahsa cohorts, respectively (P = .09). Gastrointestinal pathogens were rarely isolated in both cohorts of HCWs and acquisition of pathogens after the Hajj was documented in only a few individuals.Conclusions: HCWs providing care for pilgrims both acquire pathogens and present symptoms (especially respiratory symptoms) more frequently than those not working during Hajj.

Published
2019

32. Omega 3 and atrial fibrillation: Where are we?

Author

Annamaria Martino, Laura Pezzi, Leonardo Calò, Maria Penco, Elisa Salustri, and Roberta Magnano

Subjects
Secondary prevention, chemistry.chemical_classification, medicine.medical_specialty, Future studies, business.industry, Atrial fibrillation, Omega 3 index, 030204 cardiovascular system & hematology, Pharmacology, medicine.disease, Bioinformatics, Fish oil, Cardiac surgery, 03 medical and health sciences, 0302 clinical medicine, Editorial, chemistry, medicine, Antiarrhythmic effect, Cardiology and Cardiovascular Medicine, business, 030217 neurology & neurosurgery, Polyunsaturated fatty acid
Abstract

Anti-arrhythmic properties of n-3 polyunsaturated fatty acids, at least in part mediated by anti-oxidant, anti-inflammatory and anti-fibrotic power, have been widely proved. Effect of fish oil on atrial fibrillation, both in primary and in secondary prevention and after cardiac surgery, are controversial, mostly due to lack of homogeneity between studies but also due to individual variability in response to fatty acids administration. Inclusion of measurement of incorporation of fish oil into cell membranes, appears to be essential in future studies, to assess their antiarrhythmic effect.

Published
2016

33. Detection and Isolation of Sindbis Virus from Field Collected Mosquitoes in Timimoun, Algeria

Author

Nazli Ayhan, Aissam Hachid, Laurence Thirion, Kamel Eddine Benallal, Laura Pezzi, Fayez Ahmed Khardine, Chahrazed Benbetka, Sihem Benbetka, Zoubir Harrat, and Remi Charrel

Subjects
Infectious Diseases, Culicidae, Alphavirus Infections, Seroepidemiologic Studies, Virology, Algeria, parasitic diseases, Animals, Humans, Mosquito Vectors, Sindbis Virus, Kenya, Phylogeny
Abstract

Sindbis virus (SINV) is a zoonotic alphavirus (family Togaviridae, genus Alphavirus) that causes human diseases in Africa, Europe, Asia, and Australia. Occasionally, SINV outbreaks were reported in South Africa and northern Europe. Birds are the main amplifying hosts of SINV, while mosquitoes play the role of the primary vector. Culex mosquitoes were collected in Algeria and subsequently tested for SINV. SINV RNA was detected in 10 pools out of 40, from a total of 922 mosquitoes tested. A strain of SINV was isolated from a pool displaying high viral load. Whole-genome sequencing and phylogenetic analysis showed that the SINV Algeria isolate was most closely related to a Kenyan strain. This was the first record of SINV in Algeria and more broadly in northwestern Africa, which can be a potential risk for human health in the circulating area. Further studies are needed to measure the impact on public health through seroprevalence studies in Algeria.

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