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The Secproch working group (for "securite des produits issus du corps humain") was created in 2019 within the « Haut Conseil de la sante publique » (HCSP) for addressing all the questions related to labile blood products, organs, tissues, cells (OTC) and gametes issued from human body. It is notably in charge of the management of alerts regarding arbovirus infections. These infections due to arthropod-transmitted viruses are responsible for emergence and reemergence, notably in the context of global warming. This review relates the alerts taken into consideration by the Secproch group between 2019 and 2021 following three pathologies due to Flaviviridae : dengue, West Nile virus (WNV) infection and tick-borne encephalitis (TBE). The dengue alerts have occurred in French Indies where the virus is endemic/epidemic, Reunion Island where the population was naive until 2018 towards the virus, and the metropole where foci of autochthonous cases are observed sporadically. The WNV infection was responsible of both human and equine cases in 2019 in the South of France but with intensity much less than in 2018. At last, the TBE virus was at the origin of a cluster of about 40 cases in the Ain department following a contamination by crude non-pasteurized goat cheese. This review offers the opportunity to reevaluate the risks linked to these three viruses through blood products and organs/tissues/cells and to precise the means recommended by HCSP to secure these products.

We conducted a prospective study in healthcare workers (n=296) of the University Hospital of Lyon, France. Serum samples (n=296) collected six months after disease onset were tested using three commercial assays: the Wantai Ab assay detecting total antibodies against the receptor binding domain (RBD) of the S protein, the bioMerieux Vidas assay detecting IgG to the RBD and the Abbott Architect assay detecting IgG to the N protein. The neutralising antibody (NAb) titre was also determined for all samples with a virus neutralisation assay (VNA) using live virus. The positivity rate was 100% with the Wantai assay, 84.8% with the bioMerieux assay and 55.4% with the Abbott assay. Only 51% of HCWs were positive for the presence of NAb. Less than 10 % of HCWs had a NAb titre greater than 80. At a neutralising titre of 80, the area under the curves [IC 95%] was 0.71 [0.62-0.81], 0.75 [0.65-0.85] and 0.95 [0.92-0.97] for Wantai, Abbott and Vidas respectively. The data presented herein suggest that commercial assays detecting antibodies against the N protein must not be used in long-term seroprevalence surveys while the Wantai assay could be useful for this purpose. VNA should remain the gold standard to assess the protective antibody response, but some commercial assays could be used as first-line screening of long-term presence of NAb.

The sensitivity of NG-test CTX-M Multi assay and BL-RED test incubated 10 min for the detection of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae was 80.6% and 90.3% respectively. Using an extended 60 min incubation with the BL-RED test, its sensitivity was increased to 100% and 60.9% for ESBL-producing and cephalosporinase-overexpressing Enterobacteriaceae respectively.

This article reports the observed rate of infection with severe acute respiratory syndrome coronavirus-2 in healthcare workers (HCWs) who worked on wards dedicated to care of patients with coronavirus disease 2019 (COVID-19) compared with HCWs who worked on non-COVID-19 wards. The infection rate was significantly higher among HCWs who worked on non-COVID-19 wards (odds ratio 2.3, P=0.005), illustrating the need to strengthen social distancing measures and training.

Objectives This prospective study performed in the paediatric emergency department of the University Hospital of Saint-Etienne aimed to measure the impact of the 24/7 bedside use of the Veritor™ System (Becton Dickinson) on the reduction of supplementary investigations, hospital stay and antimicrobial use. Methods Influenza virus A and B antigens were detected with a rapid influenza digital immunoassay (DIA) on nasopharyngeal aspirates (NPAs) sampled from the children consulting at the paediatric emergency department between January and March 2016 for influenza-like illness. The same NPA was tested by immunofluorescence and/or molecular routine assays. Before performing the DIA, the clinician filled in a questionnaire listing the tests that he/she would have prescribed in the absence of the rapid testing. The prescription of complementary investigations, antimicrobial treatments and hospital stay were also compared to those of the 3 previous years. Results A total of 514 children with flu-like symptoms were included. The use of the DIA at bedside decreased the prescription of blood puncture by 47.9% (21.2% to 6.6%), of chest X-rays by 69.0% (33.3% to 10.3%), of lumbar puncture by 77.8% (7.0% to 1.6%), of urine culture by 79.2% (23.3% to 4.9%), of antibiotic treatments by 70.1% (16.9% to 5.1%), and of hospital stay by 25.0% (27.2% to 20.4%), resulting in a reduction of medical costs estimated to more than €69 000 in a season. Conclusions In addition to delivering a rapid aetiological diagnosis, this strategy saves medical costs and favours an antimicrobial stewardship strategy. However, further prospective studies are needed to confirm our findings.

The in vitro propagation of human rhinoviruses (RVs) is difficult because only few continuous human cell lines are permissive to these agents. We propose an innovative model of epithelial cell infection using a non-transformed continuous keratinocyte line from human origin (HaCaT cells). After infection with RV-A13, RV-A16 or RV-A19, HaCaT cells produced infectious particles without showing any observable cytopathic effect and overexpressed ICAM-1 (intercellular adhesion molecule 1), the major entry receptor of RVs. Furthermore, the treatment of HaCaT cells with 10 µM clarithromycin reduced the viral titer by 93% and 60% during the first and second days following viral infection, respectively, probably by down-regulating ICAM-1 expression. This original model of epithelial cell infection by RV could be useful to study chronic viral infection and bacterium-virus interactions at the cell level. These results also suggest that clarithromycin may be evaluated for treating in vivo infections associating RV to a susceptible bacterium.

A 57-year-old female experienced two successive peripheral facial paralysis (PFP) episodes following influenza immunization in 2009 and 2016 with two different vaccines. The similarity of chronology and semiology between the two events and the absence of alternative etiology plead for intrinsic accountability. Extrinsic accountability relies on previous case reports of PFP related to flu vaccination (26 cases in the French pharmacovigilance database and 4 cases in the medical literature).

Staphylococcus aureus nasal colonization is a well-known independent risk factor for infection caused by this bacterium. Screening and decolonization of carriers have been proven effective in reducing S. aureus infections in some populations. However, a gap remains between what has been proven effective and what is currently done. We aimed to summarize recommendations and current knowledge of S. aureus decolonization to answer the following questions: Why? For whom? How? When? And what are the perspectives?

Objective We aimed to assess the prevalence of Chlamydophila pneumoniae and Mycoplasma pneumoniae acute infections, using serological testing, in patients admitted to the emergency department for acute exacerbations of chronic obstructive pulmonary disease (COPD). Methods We performed a prospective observational study of 100 consecutive patients. Serum specimens were collected at day 0 and day 15. C. pneumoniae and M. pneumoniae antibodies (IgM and IgG) were tested by commercial ELISA and immunofluorescence assay, respectively. Results We did not observe any acute M. pneumoniae infection; however, 11 patients (11%) showed a profile compatible with a recent C. pneumoniae infection (nine patients with specific IgM and two with an IgG antibody rise) . Demographic and clinical parameters did not differ between patients with and without biological profile of recent C. pneumoniae infection. Conclusion C. pneumoniae is a pathogen that requires specific antimicrobial treatment. Its detection must always be performed considering its prevalence in patients presenting with acute COPD exacerbations.

Summary Objectives Legionnaires' disease (LD) is a severe disease associated with community and hospital-acquired pneumonia, frequently under diagnosed. The main aim of our study was to determine the value of PCR for the diagnosis of LD in routine clinical practice. Methods In a prospective study, from March 2007 to April 2010, the value of PCR on non-invasive respiratory specimens (NIRS) was compared to those of the other available tools for LD diagnosis in patients hospitalized for pneumonia. Results Among 254 consecutive cases of pneumonia included, 24 cases were LD (19 confirmed and 5 probable) representing the first documented microbiological etiology. Molecular diagnosis of LD was performed on NIRS by using 16S rRNA PCR, and secondarily mip PCR, with no discrepant results between the 2 methods: it was found positive in 14 cases and led to identify 2 supplementary probable cases of LD. Based on clinical and at least 2 positive LD tests, PCR yielded a better diagnostic value than antigen urinary test (12 vs 10 cases). Conclusion These results revealed that molecular diagnosis of LD on NIRS is reliable and may contribute to better identify cases of LD.

Summary Transfusion has been tainted with the risk of contracting an infection – often severe – and fears about this risk are still prevailing, in sharp contrast with the actual risk in Western countries. Those actual risks are rather immunological, technical (overload) or metabolic. Meanwhile, in developing countries and particularly in Africa, transfusion transmitted infections (TTIs) are still frequent, because of both the scarcity of volunteer blood donors and resources and the high incidence and prevalence of infections. Global safety of blood components has been declared as a goal to be attained everywhere by the World Heath Organization (WHO). However, this challenge is difficult to meet because of several intricate factors, of which the emergence of infectious agents, low income and breaches in sanitation and hygiene. This review aims at encompassing the situation of TTIs in different settings and means that can be deployed to improve the situation where this can possibly be.

Viral safety remains a major concern in transfusion of blood products. Over years, the control measures applied to blood products were made more and more sophisticated; however, the number of infectious agents, and notably of viruses, that can be transmitted by transfusion is increasing continuously. The aim of this review paper is to actualize that published in the same journal by the same authors in 2011 with more details on some of actual vs virtual viral threats that were identified recently in the field of blood transfusion. The main subjects that are covered successively concern the transmission via transfusion of hepatitis E virus, the frequency of transfusion transmitted arboviruses, transfusion at the time of the Ebola epidemics in West Africa, the debated role of Marseillevirus (giant viruses infecting amoebae and suspected to infect human blood latently), and, finally, the recent report of the identification in blood donors of a new member of the Flaviviridae family. The addition of these new viral risks to those already identified-partially controlled or not-pleads for the urgent need to move forward to considering inactivation of infectious agents in blood products.

Plasma therapy consists in bringing to a patient in need - in general suffering a severe, resistant to current therapy, and even lethal infection - plasma or specific, fractioned, antibodies, along with other immunoglobulins and possibly healing factors that can be obtained from immunized blood donors; donors (voluntary and benevolent) can be either actively immunized individuals or convalescent persons. Plasma therapy has been used since the Spanish flu in 1917-1918, and regularly then when viral epidemics threatened vulnerable populations, the last reported occurrence being the 2013-2015 Ebola virus outbreak in West Africa. The precise action mechanism of plasma therapy is not fully delineated as it may function beyond purified, neutralizing antibodies.

Blood transfusion is made possible principally by use of donated homologous components that - in turn - can be perceived as sources of danger by recipients. This may create an innate immune response dominated by inflammation, especially when transfusion is repeated. Residual leukocytes in blood components can source inflammatory lesions but considerably less than used to be prior to systematic, early and stringent - in process - leukoreduction. Every blood component can cause inflammation, though barely in the case of therapeutic plasma (in such a case, this is mainly restricted to allergy). Iron that may be freed by red blood cells but also processing and storage lesions such as the emission of microparticles can reveal themselves as pro-inflammatory. Platelets in platelet components represent the main source of inflammatory and/or allergic hazards in transfusion; this is linked with processing and storage lesions but also with the platelet physiology itself. It is of utmost importance to avoid inflammatory adverse events in patients that are fragile because of their primary condition and/or treatment; this stands for their safety, as inflammation can be extremely severe and even lethal, and also for their comfort; this increases efficacy of transfusion programs while reducing the overall costs.

From blood donor collection to transfusion of the recipient, there are several layers of protection of the blood supply. These measures combined with huge progresses over the three past decades in pathogen discovery and blood testing for specific pathogens (human immunodeficiency virus (HIV), hepatitis B (HBV) and C (HCV) viruses, Human T-cell leukemia virus (HTLV)), provide the greatest safety. With the implementation of serological and molecular testing, at least in high-income countries, transfusion-transmitted infections have become extremely rare. However, for pathogen agents, which are not tested and especially those which are responsible for emerging infectious disease, it became apparent that full control of infectious disease had not been achieved. In addition, the immune status of the recipient has also an impact in the outcome of infectious diseases transmitted by transfusion. Blood safety is based on several measures: education and deferral of donors with risk factors for transmissible disease, blood testing, pathogen reduction interventions, and patient blood management. This paper proposes a review of the residual risk of transmission of infectious diseases by transfusion and of the additional interventions able to further reduce it.

Typing of human enterovirus (EV) remains a major goal for diagnostic and epidemiological purposes. Whereas sequencing of the VP1 coding region is the reference standard for EV typing, a method relying on sequencing of the VP2 coding region has been proposed as an alternative; however, this has been validated only on cell culture supernatants. To avoid the selection of cultivable strains and to quicken the identification step, a new semi-nested PCR method targeting the VP2 region was developed by use of the CODEHOP strategy. After validation of the method on reference and clinical strains, a total of 352 clinical specimens found to be positive for EV RNA (138 with the GeneXpert EV kit and 214 with the Enterovirus R-gene kit) during a 3-year period (2010–2012) were analysed prospectively for VP2 genotyping. Overall, 204 (58%) specimens were typeable. A higher proportion of throat swab/stool specimens than of cerebrospinal fluid (CSF) specimens was found to be typeable (94 of 142 (66.2%) vs. 83 of 169 (49.1%), respectively, p

Highlights • Systematic antimicrobials are recommended in community-acquired pneumonia of child. • A large panel of bacteria and viruses was detected in 85 children exhibiting CAP. • More than 60% of children with CAP exhibited an exclusive viral infection. • A co-infection with at least 2 viruses was observed in >40% of the children. • Data suggest that the use of antimicrobials in child's CAP should be revisited., Background The management of children with community-acquired pneumonia (CAP) is largely influenced by the development of new molecular diagnostic tests that allow the simultaneous detection of a wide range of pathogens. Objectives Evaluation of a diagnostic approach including multiplex PCR assays for revisiting the epidemiology and etiology of CAP in children at hospital. Study design Children of all ages consulting at the Emergency Department of the University hospital of Saint-Etienne, France, during the 2012–2013 winter period were included. In addition to bacterial cultures, the following pathogens were detected using biplex commercially-available rt-PCR tests: adenovirus, respiratory syncytial virus, human metapneumovirus, bocavirus, rhinovirus/enterovirus, coronavirus, influenza viruses A and B, parainfluenza viruses, Mycoplasma pneumoniae and Chlamydophila pneumonia. Results From 85 patients with CAP, at least one pathogen was identified in 81 cases (95.3%), including 4 bacterial exclusive infections (4.7%), 53 viral exclusive infections (62.4%) and 24 mixed infections (28.2%). Coinfection by at least two viruses was observed in 37 cases (43.5%). Mean age was higher in the case of documented bacterial infection (P

The novel SPF10 real-time PCR assay allows the simultaneous amplification and detection of the HPV target. That way, LiPA analysis of the HPV-negative samples can be avoided, reducing workload and cost. This study aims to evaluate the performance of the SPF10 real-time PCR in combination with the LiPA assay for HPV detection and typing in cervical samples. Thirty-nine cervical samples were subjected to the SPF10 conventional PCR in combination with the LiPA assay. Subsequently, the SPF10 real-time PCR was performed to enable the comparison between the SPF10 conventional and the real-time PCR results. In case of discrepancy, the samples were subjected to the CLAR(R) HPV2 assay. As a result, 27 out of 39 samples were identified as HPV-positive by the SPF10 real-time PCR and were genotyped further by the LiPA assay. Twenty samples (74.1%) showed an absolute agreement between the conventional and real-time SPF10 PCR (concordant), three (11.1%) displayed additional or fewer types (compatible), two (7.4%) did not show any similarity between both assays (discordant) and the remaining two (7.4%) were LiPA-negative. The two assays showed an excellent strength of agreement for individual (k = 0.932) and multiple genotype detection (k = 0.834). In conclusion, the two SPF10 PCR methods are comparable. Therefore, the SPF10 real-time PCR with subsequent LiPA could be used for the detection and genotyping of HPV in cervical samples. (C) 2013 Elsevier B.V. All rights reserved.

summary Background & aims: Group B streptococci (GBS) are a major cause of neonatal infection. The existence of a link between levels of vitamin D and vaginal carriage of GBS was investigated. Methods: Vitamin D level, assayed from umbilical venous blood at birth, and the status of GBS vaginal carriage during the last term of pregnancy were available from 2246 motherechild couples. Results: Levels of vitamin D in cord blood at birth was deficient ( 30 ng/ml) in 32.1%, 53.0% and 14.9% of tested subjects, respectively. Levels differed significantly depending on the season of delivery, the duration of exposure to sunlight, the skin phototype, and on vitamin D supplementation. The percentage of GBS vaginal carriage was 15.4. A negative association was observed between GBS vaginal carriage and level of vitamin D (P < 0.01 by chi-square test). No relationship was established between GBS colonisation (27 cases identified) or infection (7 cases identified) and vitamin D status. Conclusions: A correct vitamin D level was associated with a lower rate of GBS vaginal carriage during pregnancy. This calls for the monitoring of vitamin D status in pregnant women, a screen which could greatly reduce the need for intrapartum antibiotic prophylaxis.

Resume Objectifs Resultats d’une enquete sur les vaccinations de medecins generalistes (MG) du departement de la Loire, France, en 2010. Sujets et methodes Enquete declarative par auto-questionnaire aupres de 460 MG, 288 (64 %) ont repondu. Resultats La couverture vaccinale pour DT-polio depuis moins de dix ans, BCG, coqueluche, grippe saisonniere, grippe A/H1N1 souche 2009 et hepatite B etait respectivement de 81, 74, 59, 73, 65 et 87 %. Soixante-quatre pour cent des MG ont declare etre vaccines contre la rougeole et 49 % contre la varicelle ; 76 % disaient connaitre les recommandations vaccinales des soignants, mais 41 % auraient souhaite plus d’informations sur ce theme. En analyse multivariee, les plus jeunes MG etaient mieux vaccines vis-a-vis de DT-polio, hepatite B, grippe saisonniere et rougeole. Conclusions Les vaccinations des MG (connaissances, pratiques) doivent etre ameliorees, notamment par une information annuelle, la formation continue et un suivi medical dans le cadre professionnel.

Resume Contexte Le rotavirus est le principal agent infectieux des gastroenterites aigues severes (GEA) chez le nourrisson et l’enfant de moins de cinq ans. Compte tenu de la disponibilite recente de nouveaux vaccins, il est important d’evaluer precisement le taux d’incidence des gastroenterites a rotavirus (GERV) communautaires et leurs consequences medico-epidemiologiques. Methodes Ce travail est le versant francais du programme d’etudes SPRIK. Il s’agit d’etude multicentrique, prospective, observationnelle realisee entre octobre 2005 et mai 2007, destinee a estimer chez les enfants de moins de cinq ans l’incidence annuelle des GERV qui ont conduit a une consultation chez un medecin generaliste ou un pediatre. Il presente les donnees collectees par 41 medecins generalistes et 36 pediatres repartis sur l’ensemble du territoire francais metropolitain. Un echantillon de selles a ete preleve pour chaque enfant. La presence de rotavirus a ete recherchee par le medecin a l’aide d’un test immunochromatographique rapide. Resultats Un total de 1648 episodes de GEA correspondant a 1463 patients eligibles a ete inclus dans l’etude, essentiellement de decembre a mai (pic en fevrier–mars). Le taux d’incidence des GERV donnant lieu a une consultation en medecine de ville a ete de 1357 cas pour 100 000 sujets-annees (1,36 %), avec un intervalle de confiance a 95 % de [1345–1368]. Le pic d’incidence a ete observe avant l’âge de deux ans. Les GERV ont represente 21 % de l’ensemble des GEA. Les patients ayant une GERV etaient plus jeunes (14,1 ± 10,8 mois versus 18,4 ± 13,9 mois pour les autres GEA, p p p p Conclusion Le taux d’incidence rapporte dans cette etude est proche de ceux de precedents travaux realises en Europe. La frequence et la gravite relative des GERV communautaires plaident en faveur d’une vaccination chez les tres jeunes enfants pour reduire le fardeau lie a cette pathologie.

Transfusion of labile blood products (LBPs) generates occasional inflammatory : type, hazards; for a large part of these, no antigen/antibody conflict is thus, detected. Residual leucocytes used to account for a large part of such incidents - rarely accidents. Since, however, the systematic leukoreduction of LBPs, leucocytes are the less and less incriminated in adverse events. Platelets themselves proved capable of secreting copious amounts of inflammatory mediators, even in the absence of any deliberated stimulation. Meanwhile, even though exceptionally, inflammation can be observed after red blood cell transfusion. It has been noticed that the collection mode of cellular compounds, as well as the preparation and storage conditions are capable of inflicting lesions to the cell membranes and to activate those cells, and thus promoting inflammatory responses. Storage lesions as well as ageing of the stored cells alongside with cell apoptosis contribute to inflammatory responses. This present 'State of the Art' paper aims at encompassing the primary and secondary components of the LBPs, along with the various types of molecules displaying pro-inflammatory properties that can be encountered in transfusion. A better knowledge of causes of inflammatory transfusion-linked hazards is indeed instrumental to the implementation of safety measures aimed at reducing or suppressing these unwanted effects.

Platelets are currently acknowledged as cells of innate immunity and inflammation and play a complex role in sepsis. We examined whether different types of LPS have different effects on the release of soluble signaling/effective molecules from platelets. We used platelet-rich plasma from healthy volunteers and LPS from two strains of gram-negative bacteria with disparate LPS structures. We combined LPS-stimulated platelet supernatants with reporter cells and measured the PBMC cytokine secretion profiles. Upon stimulation of platelets with both Escherichia coli O111 and Salmonella minnesota LPS, the platelet LPS::TLR4 interaction activated pathways to trigger the production of a large number of molecules. The different platelet supernatants caused differential PBMC secretion of IL-6, TNFα, and IL-8. Our data demonstrate that platelets have the capacity to sense external signals differentially through a single type of pathogen recognition receptor and adjust the innate immune response appropriately for pathogens exhibiting different types of 'danger' signals.

Resume Contexte En France, les infections a Rotavirus sont responsables chaque annee d’environ 300 000 episodes de gastroenterite aigue (GEA) chez les enfants de moins de cinq ans. Elles sont egalement a l’origine de 138 000 consultations en ville et de 18 000 hospitalisations. L’etude Surveillance for Hospitalized Rotavirus Infections in Kids (SHRIK) est une etude prospective observationnelle europeenne realisee afin d’evaluer le fardeau des gastroenterites a Rotavirus (GERV) chez ces enfants. Methode L’etude SHRIK se deroulait sur une periode d’un an et s’adressait aux patients de moins de cinq ans ayant consulte le service des urgences pour une gastroenterite aigue (GE-U), ayant ete hospitalises pour une gastroenterite aigue (GE-H) ou ayant developpe une GERV 48 heures apres leur hospitalisation (GE-N). Un echantillon de selles etait preleve pour chacun des enfants inclus et analyse par methode Elisa. Resultats Les resultats presentes concernent les patients inclus en France. Un total de 755 patients atteints de GEA et âges de moins de cinq ans a ete inclus (357 pour une GE-U, 372 pour une GE-H et 26 pour une GE-N). Parmi ces patients, la proportion de GERV etait de 49,1 % pour les GE-U (n = 114) et de 64,4 % pour les GE-H (n = 186). La majorite (89 %) des GERV concernait des enfants de moins de deux ans. Selon l’echelle de Vesikari, la GEA des patients GERV etait plus frequemment qualifiee de « severe » que celle des patients atteints de gastroenterite non a Rotavirus (GENRV) (68,4 % contre 41,9 %, p Conclusion L’etude SHRIK montre que le fardeau des infections a Rotavirus est eleve ; elles representent plus de 70 % des GEA traitees a l’hopital au cours du pic hivernal.

During the last 20 years, the safety of blood products increased dramatically with regard to the infectious risk and notably to that represented by retroviruses (HIV and HTLV) and hepatitis B and C viruses. The aim of this review is to identify the residual and emergent viral threats that could be responsible for the occurring of new contaminations in the receivers of blood products. Beside many other viruses (HHV-8, erythrovirus B19, hepatitis A and E viruses...), a special attention has been paid to emerging arbovirus diseases (West Nile virus infection, dengue, chikungunya) that threaten to occur in the French metropolitan area following the implantation in Europe of the mosquito Aedes albopictus, the main vector of dengue and chikungunya in temperate regions. Another blood-linked risk, notably in United Kingdom and France, is the prion agent responsible for the variant form of the Creutzfeldt-Jakob disease. The review is concluded by a brief overview of the measures aimed to control these emergences, including the exclusion of at-risk donors, the diagnostic tests able to detect a specific agent, the leukocyte reduction of labile blood products, and the physical or chemical treatments aiming the nonspecific inactivation of infectious agents potentially present in blood without impairing significantly the physiological properties of blood compounds. The ability to control prospectively the new viral risks linked to blood products is a challenge for the preservation of the confidence of both clinicians and receivers in the safety of blood transfusion.

The risk factors for Staphylococcus aureus nasal carriage have been mainly defined in healthy volunteers. Using data from a large multicenter prospective clinical cohort of orthopedic surgical patients we showed that obesity (BMI >30) and younger age were independent risk factors for persistent carriage. In the absence of recommendations for S aureus decolonization in orthopedic surgery, this study suggests that at least obese patients should be targeted to prevent surgical site infections.

Resume Les microbicides peuvent se definir comme des substances topiques susceptibles de reduire le risque d’infections sexuellement transmissibles. L’objet de cette revue est de presenter un etat des lieux des connaissances relatives aux microbicides vaginaux susceptibles de proteger les femmes contre l’infection par le virus de l’immunodeficience humaine de type 1 (VIH-1). Les molecules microbicides a visee anti-VIH comprennent des composes non specifiques comme les detergents, les polyanions ou les inhibiteurs d’entree, et des molecules specifiques de VIH-1 qui incluent principalement des inhibiteurs des etapes precoces de l’infection virale et des inhibiteurs de la reverse transcriptase. Bien que l’approche microbicide soit tres seduisante sous l’angle conceptuel, aucun essai clinique controle et randomise, conduit avec des molecules de premiere generation n’a fait la preuve de l’efficacite preventive de ces composes contre l’infection par VIH-1. Des resultats encourageants ont ete obtenus chez des primates non humains et plusieurs essais cliniques menes a travers le monde devraient livrer leur verdict assez vite. Cette revue discute ces differents aspects et encore l’evaluation preclinique de la balance benefices-risques des produits microbicides, les associations de composes microbicides anti-VIH-1 et la capacite de ces substances a selectionner des mutants resistants. Une meilleure connaissance des modes de franchissement de la muqueuse cervico-vaginale par VIH-1 devrait permettre d’affiner les strategies microbicides qui, en l’absence de vaccination preventive a moyen terme, constituent une alternative a evaluer pour proteger les femmes – et notamment les plus vulnerables d’entre elles – vis-a-vis de l’infection par VIH-1.

In this study, we evaluated the potential direct role of enterovirus (EV) cardiac infections in the pathogenesis of myocardial infarction (MI).Enteroviruses (Picornaviridae) have been suspected to play a role in the development of acute MI.The presence of EV ribonucleic acid (RNA) sequences and capsid viral protein 1 (VP1) and the virus-mediated focal disruption of dystrophin were retrospectively investigated by reverse transcriptase-polymerase chain reaction and immunohistochemistry assays in endomyocardial tissues of patients who died suddenly of acute MI by comparison with similar samples of control patients matched for gender, residence area, and year of death.Enterovirus infection markers were detected in 20 (40%) of 50 patients who died suddenly of MI, 2 (4%) of 50 matched subjects without cardiac disease (p0.001), and 4 (8%) of 50 matched patients exhibiting a noncoronary chronic cardiopathy (p0.001). All of the EV RNA-positive patients exhibited VP1, which provided evidence of viral protein synthesis activity. The VP1 gene sequences amplified after cloning from myocardial or coronary samples of 8 of the MI patients and showed a strong homology with sequences of coxsackievirus B2 and B3 serotypes. Moreover, in the endomyocardial tissue of these 8 patients, immunohistochemical analyses demonstrated that there was disruption of the sarcolemmal localization of dystrophin in the same tissue areas that were infected by coxsackieviruses.Our findings demonstrate a significantly higher proportion of active coxsackievirus B cardiovascular infections in patients who suddenly died of MI compared with matched control subjects, suggesting that these EVs may significantly contribute to the pathogenesis of acute MI by a focal disruption of the dystrophin-glycoprotein complex.

Background Various pathogens have been suspected to play a role in the initiation or amplification of the atherosclerotic lesions. Both experimental and epidemiological arguments plead for a possible role of enterovirus in this process. Objective To determine the prevalence of enterovirus genome in atherosclerotic plaques, in comparison with Chlamydia pneumoniae, human cytomegalovirus (hCMV) and herpes simplex virus. Study design Pilot study on 18 patients who underwent artery resection. Five artery samples were tested for each patient and each pathogen by using PCR techniques whose sensitivity was evaluated for this kind of specimen. The quality of the extraction step was assessed by amplification of a fragment of the human aldolase A gene. Results The genome of at least one infectious agent was detected in artery samples from 7 of the 18 patients (38.9%). In all cases, only one of the five aliquots was found positive; a confirmation was done by sequencing the PCR product. With regards to enterovirus, four patients (22.2%) were detected positive (one of them being also positive for hCMV). Conclusions These results suggest that small amounts of enterovirus genome are commonly found in lesions of patients with advanced arteriosclerosis. Further studies are needed to evaluate the clinical significance of this association.

Objective. – Most of coxsackieviruses A (CV-A) are difficult to isolate in cell culture and are responsible for flask paralysis in suckling mice. The aim of the present work was to analyze the ability of immune and RT-PCR techniques to detect viral components of three different serotypes, CV-A6, CV-A13, and CV-A14, in skeletal muscles of experimentally infected suckling mice. Material and methods. – The antigen detection was done by immunofluorescence technique on trypsinized muscular cells and by immunoperoxidase assay on frozen sections of skeletal muscle, using a monoclonal antibody directed towards a conserved epitope of the VP1 capsid protein among enteroviruses. The nested RT-PCR technique used primers located in the 5′ non coding region of viral RNA. Results. – The group antigen was present in muscle cells of suckling mice infected by the three serotypes of CV-A which were assayed. Similarly, the muscle specimens were positive by nested RT-PCR. A kinetic study performed with CV-A13 and CV-A14 showed that the RT-PCR assay was positive as soon as 24 h after infection whereas the detection of VP1 antigen and symptoms of flask paralysis were observed only 48 and 72 h after infection, respectively. Conclusion. – These results show that the tested serotypes of CV-A can be easily detected in muscle specimens of suckling mice by using antigenic and molecular techniques currently available for the diagnosis of enterovirus infections.

Non-fermentative Gram negative rods are opportunistic pathogens responsible for nosocomial infections. Using phenotypic markers (serotypes for Pseudomonas aeruginosa and antibiotic susceptibility) allows a preliminary screening of epidemiologically-related strains. However, genotypic markers are necessary to better characterize nosocomial strains for the investigation of outbreaks or cross-transmissions in the hospital setting. Infections due to P. aeruginosa, Burkholderia. cepacia or Stenotrophomonas. maltophilia are usually hospital-acquired and responsible for a high mortality rate as illustrated by the lethality of nosocomial pneumonia due to P. aeruginosa. The severity of these infections is due to the virulence factors of the bacteria and to their occurrence in debilitated patients in whom invasives devices are used. The hospital environment can act as a reservoir with a rate of exogeneous transmission of these bacteria as high as 50% in some studies. To better prevent nosocomial infections related to Gram negative non fermentative rods, the control of the aqueous hospital environment, the strict application of hand disinfection and the investigation of potential cross-transmission in the hospital setting are needed.

Resume Objectif. – Etudier la distribution des differents sous-types de HIV-1 dans la population de sujets depistes positifs pour ce virus au CHU de Saint-Etienne, France, et decrire les caracteristiques epidemiologiques de ceux infectes par un sous-type non B. Methodes. – Enquete epidemiologique portant sur 271 sujets infectes par HIV-1 suivis dans le service de maladies infectieuses du CHU de Saint-Etienne sur une periode de 20 ans. Les echantillons ont tous beneficie d'un serotypage et ont ete en partie genotypes. Resultats. – Deux cent soixante-quatre sujets ont ete depistes comme porteurs d'une souche HIV-1 de groupe M, dont 191 hommes et 73 femmes. En combinant les donnees du serotypage et du genotypage, il a ete identifie 195 sujets porteurs de souches B et 69 porteurs de souches non B. Parmi ces dernieres, sont majoritaires le sous-type A et des formes recombinantes apparentees. L'origine non francaise du sujet, le mode de transmission heterosexuel et le sexe feminin sont statistiquement relies au portage d'une souche non B. L'incidence dans le temps des souches non B croit progressivement (pour depasser 40 % actuellement) ; cette progression est particulierement sensible chez les hommes nes en France ayant des pratiques sexuelles a risque. Conclusion. – Ces resultats soulignent que plus de 40 % des nouveaux cas d'infections par HIV-1 recenses dans la region stephanoise sont en relation avec des souches de sous-types non B et que des souches de genotypes A et apparentes circulent desormais au sein de la population autochtone exposee a des pratiques sexuelles a risque.

The evaluation of the fully automated BD MAX Cdiff assay on a panel of 100 stool samples characterized by the Xpert C. difficile assay reported a high concordance between the two molecular assays (kappa coefficient of 0.96), which makes this new assay suitable for routine detection of toxigenic Clostridium difficile.

Hemodialysed patients are recognised as a group at increased risk of infection with hepatitis C virus (HCV). The aim of this study was to determine the prevalence and incidence of HCV infection among dialysis patients of the east-centre part of Tunisia. Two hundred and seventy-six patients dialysed until 2001 were recruited within seven hemodialysis units located in the cities of Sousse, Monastir and Mahdia. The serum markers of HCV infection were tested over the period of March 2000–December 2002, by a 3rd generation ELISA test for antibodies and by qualitative RT-PCR technique for viral RNA. The prevalence of anti-HCV antibodies and of HCV RNA was 32.6% (90 patients) and 25.7% (71 patients), respectively. Between 1998 and 2002, 20 new infections were documented in five of the seven dialysis units corresponding to an incidence of 2.34% per year, with an average time of contamination after the beginning of dialysis of 4.6 years. If all the infections are assessed to have occurred during dialysis, the density of incidence of HCV contamination was 4.4% per year of dialysis. A high correlation was noticed between the presence of HCV markers in serum and the duration of dialysis (F = 34.15, P < 0.0001). In the absence of other risk factors (transfusion, drug-addiction), these results plead for the nosocomial transmission of the observed HCV infections. A phylogenetic analysis of the E2 hypervariable region of the viral genome is in progress to confirm this assumption.

The aim of this study was to evaluate the risk of hospital-acquired diarrhoea during an epidemic period through a prospective multicentre observational study. A systemic investigation of the hospital-acquired diarrhoea (occurring at least 48 h after hospital admission) was conducted through a standardised questionnaire from January to March 1999 in patients of 5 years old or less hospitalised in 28 wards (620 beds) belonging to 20 hospitals located in the south-east part of France. Overall, 241 cases of hospital-acquired diarrhoea were collected, corresponding to a prevalence of 3.3% (3.6% after exclusion of patients admitted for diarrhoea) and a density of incidence of 0.81 per 100 days of hospitalisation. The mean stay duration of hospital-infected patients was greater than 10 days, versus 3.9 days for the other children (P < 0.001). A readmission was required in 27% of the infected children. Rotavirus was involved in 97.8% of microbiologically documented cases (88%). In 50% of the cases, the hospital-acquired diarrhoea was seen in patients with bronchiolitis. Contact isolation measures were prescribed in 88.4% of the cases. These results stress that hospital-acquired diarrhoea represent an important medical and economic load for paediatric units and could be used as reference data to evaluate the impact of preventive measures, especially to reduce readmission and mean stay duration.

We report an outbreak of infections due to methicillin-resistant Staphylococcus aureus (MRSA) in a medical unit and the possible implication of student nurses in the dissemination of the epidemic strain. A retrospective epidemiological study looking for hospitalised patients colonised or infected with MRSA from the 1st of June to the 30th of September 2001 in the unit was conducted. An audit of delivered cares and a nasal screening of health care workers (HCW) was performed. Six patients were colonised or infected with a MRSA strain, four of them exhibiting a bacteremia. Six HCW had a nasal carriage of MRSA. Typing of the MRSA strains by pulsed field gel electrophoresis demonstrated an epidemic clone isolated from five of six patients, two student nurses and one HCW not implicated in nursing cares. This report illustrates the risk of nosocomial outbreak linked to cares delivered by student nurses.

Resume Ce travail decrit deux phenomenes epidemiques a Staphylococcus aureus de sensibilite diminuee aux glycopeptides, l’un en 2000 avec huit patients concernes et l’autre en 2001–2002 avec 16 patients. Le depistage systematique de ces souches aux laboratoires de bacteriologie de notre hopital a permis de les detecter rapidement. Le caractere clonal de ces souches a ete confirme par electrophorese en champ pulse. La gestion de ces phenomenes epidemiques a confirme (i) la necessite de l’application systematique des precautions standards, (ii) l’importance de la transmission d’informations pour lutter contre les bacteries multiresistantes mais egalement la difficulte de transfert dans un autre service des patients colonises et (iii) le caractere intermittent du portage de S. aureus, ce qui necessite une surveillance prolongee des patients colonises et/ou infectes. De plus cette etude souligne l’interet d’une prise en charge multidisciplinaire de la diffusion des bacteries multiresistantes.

Objective To investigate the presence of the genome of GB virus C/hepatitis G virus (GBV-C/HGV) in semen and saliva from HIV-1-infected men. Methods Samples of blood from 33 men seropositive for HIV-1 were tested for the presence of GBV-C/HGV markers of infection, RNA by RT-PCR, and anti-E2 antibodies by ELISA, respectively. The cell-free fractions of seminal fluid and saliva samples of the patients with positive blood samples for GBV-C/HGV RNA or anti-E2 antibodies were then analyzed for the presence of the RNA of this virus. In addition, six semen samples and 11 saliva samples from GBV-C/HGV-negative men were tested. Results The GBV-C/HGV RNA tested by RT-PCR was recovered from blood in 11 patients of 33 (33.3%), and the antibodies to E2 envelope protein were detected in six patients (18.2%). Since no patient was positive for both markers, the overall prevalence of GBV-C/HGV infection was 51.5% in the studied population. Four—all belonging to the homosexual risk group—of the 17 men with markers to GBV-C/HGV in blood were found to be positive for GBV-C/HGV RNA in mucosal samples: two of them exhibited genomic RNA in both semen and saliva, and two others were positive for semen only. The absence of inhibitors of the PCR technique was confirmed in all mucosal fractions found negative for GBV-C/HGV RNA, except for one saliva sample and one seminal fluid sample. Conclusion These results confirm the high prevalence of GBV-C/HGV infection in patients infected with HIV-1 by sexual exposure and the presence of GBV-C/HGV RNA in seminal fluid and saliva of men with markers of this virus in the blood, suggesting that mucosal fluids could be a potential source for the spread of the GBV-C/HGV infection.

Introduction La legionellose est une pathologie grave associee a une mortalite elevee. Mieux diagnostiquer cette pathologie est essentiel afin d’optimiser le traitement antibiotique. L’objectif de cette etude etait de mesurer la frequence de cette infection au sein d’une cohorte de patients hospitalises pour pneumopathie. Materiels et methodes Etude prospective monocentrique etudiant les caracteristiques des patients adultes hospitalises pour pneumopathie communautaire ou nosocomiale (hors pneumopathies acquises sous ventilation mecanique). La recherche de Legionella spp. a ete realisee systematiquement par culture, antigenurie, PCR ARN 16S sur crachats et serologie. Les autres etiologies infectieuses ont ete recherchees par culture, serodiagnostic (Mycoplasma pneumonia et Chlamydophila pneumoniae) ou PCR (cytomegalovirus et bacteries atypiques); les virus respiratoires courants n’ont pas ete recherches. Resultats De mars 2007 a avril 2010, 254 patients ont ete inclus dont 156 hommes (âge moyen: 61,3 ans). Le delai entre la survenue des symptomes et l’admission etait de 8,4 jours; 61,2 % des patients avaient recu un antibiotique avant l’admission a l’hopital. Une exposition a risque de legionellose dans les 15 jours precedents le debut des signes a ete retrouvee chez 32 % des patients (78/244). Il s’agissait d’une pneumopathie nosocomiale dans 12,2 % des cas. L’imagerie pulmonaire a mis en evidence une pneumopathie bilaterale dans 35,6 % des cas et systematisee dans 64,1 % des cas. Seulement 4,3 % des patients ont ete hospitalises en reanimation. Au sein des 86 agents infectieux detectes, on denombre 24 cas (28 %) d’infection a Legionella pneumophila (19 cas confirmes, 5 cas probables), 36 pneumonies a germe pyogene (dont 17 pneumocoques et 9 staphylocoques dores), 23 pneumonies a germe atypique (dont 18 M. pneumoniae et 5 C. pneumoniae), une tuberculose, une nocardiose et une infection a cytomegalovirus. La PCR legionelle avait respectivement une sensibilite et specificite de 58,3 % et 100 %, comparees a 66,6 % et 100 % pour l’antigenurie, 29,2 % et 100 % pour la culture et 37,5 % et 100 % pour la serologie. Une guerison ou amelioration a ete observee chez 94,3 % des patients au cours des 30 jours de suivi. Comparativement aux autres patients, les cas de legionellose etaient plus souvent tabagiques ( p p p p Conclusion Cette etude montre que Legionella spp. est une cause frequente de pneumopathie, peu distincte cliniquement par rapport aux pneumopathies liees a d’autres causes. La PCR sur crachats ameliore la detection de cette infection et devrait etre utilisee plus largement afin d’optimiser la prise en charge de cette pathologie.

Resume Objectif Le tetanos est une pathologie du sujet âge dans les pays developpes. Les auteurs ont etudie l'immunite humorale de patientes âgees de facon prospective sur une periode de 42 mois. Patientes et methodes Cent soixante femmes âgees (âge moyen: 84,5 ans), entrant en service de long sejour, ont recu un schema vaccinal comportant deux injections a un mois d'intervalle et un rappel a un an. L'immunite humorale a ete appreciee par dosage des anticorps antitetaniques par hemagglutination passive. Resultats La tolerance de la vaccination a ete tres bonne chez tous les patients. La protection prevaccinale etait satisfaisante chez 8 % des sujets, insuffisante chez 10 % et absente dans 82 % des cas. La reponse a la vaccination a ete globalement satisfaisante, puisque apres le rappel a un an, plus de 90 % des sujets non immuns apres les deux premieres injections ont fait une seroconversion. Cependant, la reponse humorale a montre une rapide decroissance chez plus de la moitie des patients survivants dans un delai de 30 a 42 mois. Des facteurs en relation avec une baisse de l'immunite cellulaire, comme l'absence de reponse a l'intra-dermoreaction a la tuberculine ou la presence d'une pathologie cancereuse, etaient associes de facon statistiquement significative a une baisse de la reponse humorale postvaccinale. Conclusion L'ensemble de ces resultats plaident pour un renforcement de la vaccination antitetanique chez les sujets âges, avec un rapprochement des injections vaccinales de rappel.

Background: Mab 5-D8/1 is a monoclonal antibody (Mab) that was shown to be directed towards a conserved epitope of the capsid protein VP1 among the genus enterovirus. The use of this Mab for the routine detection of enteroviruses in clinical specimens led to the observation that several strains of echovirus type 11 (EV-11) could not be detected on spontaneously detached cells from 26-h cultures using a two-step immunofluorescence (IF) assay. Conversely, these strains were detected positive with the same Mab when tested on adherent or trypsinizated cells. Objectives: The aim of this study was to understand the misrecognition of some strains of EV-11 by this Mab. Study design and results: IF tests at different times of the viral cycle brought evidence that the detection of a variant strain of EV-11 decreased rapidly with time, becoming undetectable 26 h post-infection, since the reference strain remained positive up to 46 h post-infection. The infective titres of the variant strains were shown to be high in comparison with those of well-recognised strains. Sequencing the Mab binding epitope confirmed that the variant strains exhibited no antigenic shift. Conclusion: These results suggest that the poor recognition of some strains of EV-11 by Mab 5-D8/1 is due to a rapid decrease of the expression of the binding epitope in the cell, maybe in relation with the high lytic power of these strains. From a practical point of view, our data indicate that a negative result when Mab 5-D8/1 is used for enterovirus typing must be interpreted cautiously with highly replicative strains and that detached cells should not be used for enterovirus identification under these circumstances.

Background: Previous studies have reported the role of enteroviruses in chronic diseases, using in-house RT-PCR protocols. A well-standardized PCR assay (Amplicor enterovirus, Produits Roche) designed for the diagnosis of enterovirus meningitis in cerebrospinal fluids (CSF) was recently described. Objectives: To evaluate this commercially-available PCR assay for the detection of enterovirus in intestinal biopsies. Study design: In order to obtain large quantities of infected material, eight mice were inoculated orally with 2×105 50% tissue culture infective doses (TCID50) of coxsackievirus B3 (CBV3); two mice were sacrificed every day from day 1 to day 4 post-infection. Stool specimens and small bowel fragments were taken from infected animals and controls. Four protocols of RNA extraction from intestinal tissue were compared. Extracted RNA was then tested by the Amplicor assay and by a seminested in-house PCR. Results: The best results were obtained with a commercial reagent using a combination of guanidium thiocyanate and phenol (TRI Reagent, Sigma). This procedure allowed the detection of enteroviral RNA in intestinal samples of 7/8 and 8/8 infected mice by Amplicor assay and seminested PCR, respectively, whereas only five samples were tested positive by conventional cell culture. When tested on serial dilutions of CBV3 mixed with intestinal tissue, a sensitivity of 0.2 TCID50/mg was achieved with both PCR assays. Conclusions: The data demonstrate that the Amplicor enterovirus assay, which is designed to avoid false-positive amplifications, can be used, with a slight modification of the RNA extraction step, for the detection of enterovirus in specimens different from CSF such as intestinal tissue.

Fourteen serotypes are currently recognized in the Ureaplasma urealyticum species. These serotypes have been divided into two genomic clusters or biovars by a large number of typing methods. The parvo-biovar includes strains of serotypes 1, 3, 6 and 14 and the T960-biovar, strains belonging to the ten other serotypes. In this study, arbitrarily primed polymerase chain reaction (AP-PCR) has been applied to the analysis of reference strains of the 14 U. urealyticum serotypes. By using two different sets of 10-mer oligonucleotide primers, the method allowed the clear differentiation between the two known biovars of the species. However, further differentiation within a same biovar was only achieved for a few standard strains of the T960-biovar analysed by using a pairwise combination of primers. The reproducibility of AP-PCR profiles was shown on strains tested after repeated subcultures and with different thermal cyclers. Additional experiments were performed on forty isolates of U. urealyticum recovered from subjects of various origins. They confirmed that AP-PCR was able to identify the strains at the biovar level. With reference to the other typing methods, AP-PCR is easy to perform and can be applied to large numbers of strains for epidemiological purposes.