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1. Differential Performance of the FilmArray Meningitis/Encephalitis Assay To Detect Bacterial and Viral Pathogens in Both Pediatric and Adult Populations

Author

Aurélie Schnuriger, Sophie Vimont, Alexandre Godmer, Joël Gozlan, Salah Gallah, Muriel Macé, Valérie Lalande, Kenda Saloum, Marine Perrier, Nicolas Veziris, and Laurence Morand-Joubert

Subjects
meningitis/encephalitis, pediatric/adult population, performance, sensitivity/specificity, syndromic assay, Microbiology, QR1-502
Abstract

ABSTRACT Meningitis/encephalitis (ME) syndromic diagnostic assays can be applied for the rapid one-step detection of the most common pathogens in cerebrospinal fluid (CSF). However, the comprehensive performance of multiplex assays is still under evaluation. In our multisite university hospital of eastern Paris, France, ME syndromic testing has been gradually implemented since 2017 for patients with neurological symptoms presenting to an adult or pediatric emergency unit. We analyzed the results from the BioFire FilmArray ME panel versus standard routine bacteriology and virology techniques, together with CSF cytology and clinical data, over a 2.5-year period to compare the diagnostic accuracy of the FilmArray ME panel to that of the reference methods. In total, 1,744 CSF samples from 1,334 pediatric and 336 adult patients were analyzed. False-positive (mostly bacterial) and false-negative (mostly viral) cases were deciphered with the help of clinical data. The performance of the FilmArray ME panel in our study was better for bacterial detection (specificity >99%, sensitivity 100%) than viral detection (specificity >99%, sensitivity 75% for herpes simplex virus 1 [HSV-1] and 89% for enterovirus), our study being one of the largest, to date, concerning enteroviruses. The use of a threshold of 10 leukocytes/mm3 considerably increased the positive agreement between the results of the FilmArray ME panel and the clinical features, especially for bacterial pathogens, for which agreement increased from 58% to 87%, avoiding two-thirds of inappropriate testing. Based on this analysis, we propose an algorithm for the use of both syndromic and specific assays for the optimal management of suspected meningitis/encephalitis in adult and pediatric patients. IMPORTANCE Based on our comparative analysis of performances of the diagnostic assays, we propose an algorithm for the use of both syndromic and specific assays, for an optimal care of the meningitis/encephalitis threat in adult and pediatric patients.

Published
2022
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2. Rapid detection of Clostridium difficile toxin A in stool specimens

Author

Muriel Macé, Jean-Claude Petit, Frédéric Barbut, Patrick Tilleul, and Valérie Lalande

Subjects
Microbiology (medical), toxin A, medicine.diagnostic_test, diagnosis, diarrhea, Clostridium difficile toxin A, Clostridium difficile, General Medicine, Biology, Rapid detection, Microbiology, Diarrhea, Infectious Diseases, Immunoassay, medicine, medicine.symptom, Clinical record
Abstract

OBJECTIVE: To evaluate a rapid (15-min) enzyme immunoassay in the format of an individual cassette (ImmunoCard toxin A, Meridian, BMD, Marne-la-Vallée, France) for the detection of Clostridium difficile toxin A in stool specimens. METHODS: We compared this new test with the cytotoxicity assay using MRC-5 cells, the ToxA test (TechLab, BioWhittaker, Fontenay-sous-bois, France) and toxigenic culture for the diagnosis of C. difficile-associated diseases (CDAD). A total of 236 stool specimens collected from 220 patients was simultaneously tested with the four methods. Discordant results were resolved by reviewing patients' clinical records. RESULTS: The prevalence of CDAD was 13.9%. Test sensitivities and specificities were 100% and 99% respectively for the cytotoxicity assay, 87.5% and 100% for ImmunoCard toxin A, 77.4% and 100% for the ToxA test and 100% and 98% for toxigenic culture. CONCLUSIONS: The ImmunoCard Toxin A is a very rapid, individual and easy-to-perform test for the diagnosis of CDAD. It provides same-day results and may be useful for both guiding appropriate treatment and controlling nosocomial spread of C. difficile.

Published
1997

3. Diagnostic Value of Reverse Transcription-PCR of Amniotic Fluid for Prenatal Diagnosis of Congenital Rubella Infection in Pregnant Women with Confirmed Primary Rubella Infection

Author

Isabelle Parent du Châtelet, Liliane Grangeot-Keros, C Six, Denis Cointe, Didier Ingrand, Muriel Macé, and Daniel Lévy-Bruhl

Subjects
Microbiology (medical), medicine.medical_specialty, Amniotic fluid, Rubella Syndrome, Congenital, Prenatal diagnosis, Sensitivity and Specificity, Rubella, Virus, Congenital Rubella, Pregnancy, Prenatal Diagnosis, Virology, Humans, Medicine, Pregnancy Complications, Infectious, Reverse Transcriptase Polymerase Chain Reaction, business.industry, Obstetrics, Infant, Newborn, Amniotic Fluid, medicine.disease, Fetal Diseases, Rubella Infection, Immunology, Female, Viral disease, business, Rubella virus
Abstract

The diagnostic value of RT-PCR of amniotic fluid (AF) for prenatal diagnosis of congenital rubella virus infection in 45 pregnant women with confirmed primary infection was assessed. Specificity of RT-PCR was 100% and sensitivity ranged between 83 and 95%.

Published
2004

4. Outbreak of Shiga Toxin-Producing Escherichia coli O104:H4 Associated With Organic Fenugreek Sprouts, France, June 2011

Author

Benoît Vendrely, Nathalie Jourdan-DaSilva, François Terrier, G Delmas, Edouard Bingen, P. Rolland, Dieter Van Cauteren, Elisabeth Couturier, Martine Charron, Henriette de Valk, Yahsou Delmas, Nathalie Ong, Muriel Macé, Francisco Nogareda, François-Xavier Weill, Patricia Mariani-Kurkdjian, Véronique Vaillant, Christian Combe, G. Gault, Harold Noel, Estelle Loukiadis, C Castor, Véronique Goulet, Delphine Thevenot, L. A. King, Yann Le Strat, B. Aldabe, Département des maladies infectieuses, Institut de Veille Sanitaire (INVS), European Programme for Intervention Epidemiology Training ( EPIET ), European Centre for Disease Prevention and Control ( ECDC ), Centre National de Référence des Escherichia coli, Shigella et Salmonella - Bactéries pathogènes entériques ( CNR-ESS ), Institut Pasteur [Paris], Service de Microbiologie [R Debré], Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Robert Debré-Université Paris Diderot - Paris 7 ( UPD7 ) -PRES Sorbonne Paris Cité, VetAgro Sup ( VAS ), Cellule INVS Région Aquitaine, CHU Bordeaux [Bordeaux], Hôpital d'Instruction des Armées Robert Picqué, Service de Santé des Armées, The Institut de Veille Sanitaire is supported by public funds allocated by the French Ministry of Health. The Cellule de l’Institut de Veille Sanitaire en Région Aquitaine is supported by public funds allocated by the French Ministry of Health. The Laboratoire associé au Centre national de référence des Escherichia coli et Shigella at the Hôpital Robert Debré is supported by public funds allocated by the French Ministry of Health. The Centre national de référence des Escherichia coli et Shigella at the Pasteur Institut is supported by funds from the French Ministry of Health and from the Pasteur Institut. VetAgro Sup, Campus vétérinaire de Lyon, Laboratoire d’études des microorganismes alimentaires pathogènes is supported by public funds from the French Ministry of Agriculture, Food, Fisheries, Rural Environments and Territory and the French Ministry of Finance, Industry and the Economy. The Centre Hospitalier Universitaire Bordeaux is supported by public funds allocated by the French social welfare system. The Hôpital d’instruction des armées Robert Picqué is supported by public funds allocated by the French social welfare system., European Programme for Intervention Epidemiology Training (EPIET), European Centre for Disease Prevention and Control (ECDC), Centre National de Référence - National Reference Center Escherichia coli, Shigella et Salmonella (CNR-ESS), AP-HP Hôpital universitaire Robert-Debré [Paris], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS), European Centre for Disease Prevention and Control [Stockholm, Sweden] (ECDC), and Institut Pasteur [Paris] (IP)

Subjects
Microbiology (medical), medicine.medical_specialty, Attack rate, [ SDV.MP.BAC ] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, Microbiology, 03 medical and health sciences, symbols.namesake, Escherichia coli O104:H4, Internal medicine, medicine, Poisson regression, 030304 developmental biology, 0303 health sciences, 030306 microbiology, business.industry, Outbreak, [ SDV.SPEE ] Life Sciences [q-bio]/Santé publique et épidémiologie, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, 3. Good health, Diarrhea, Infectious Diseases, Relative risk, symbols, Bloody diarrhea, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, medicine.symptom, business, Cohort study
Abstract

International audience; BACKGROUND:On 22 June 2011, 8 patients with hemolytic uremic syndrome (HUS) or bloody diarrhea were reported in France. All 8 were attendees of a community center event on 8 June near Bordeaux. Three Escherichia coli cases were confirmed by isolation of Shiga toxin-producing E. coli O104:H4 stx2 aggR producing a cefotaximase (CTX-M) β-lactamase (STEC O104:H4); the same rare serotype caused the outbreak in Germany in May-July 2011. An investigation was initiated to describe the outbreak, identify the vehicle for infection, and guide control measures.METHODS:We conducted a retrospective cohort study among all adults attending the event, including food handlers. A standardized questionnaire was administered to participants. A case was an attendee who developed HUS or diarrhea between 8 and 24 June. Cases were confirmed by isolation of STEC O104:H4 or O104 serology. Relative risks (RRs) and 95% confidence intervals (CIs) by exposure were calculated using a Poisson regression model.RESULTS:Twenty-four cases were identified (14% attack rate). Of these, 18 (75%) were women, 22 (92%) were adults, 7 (29%) developed HUS, 5 (21%) developed bloody diarrhea, and 12 (50%) developed diarrhea. Ten (42%) cases were confirmed. Fenugreek was the only sprout type with an independent association to illness (RR, 5.1; 95% CI, 2.3-11.1) in multivariable analysis.CONCLUSIONS:This investigation identified a point-source STEC O104:H4 outbreak associated with consumption of fenugreek sprouts. Comparison of results from French and German STEC O104:H4 outbreak investigations enabled identification of a common food vehicle, fenugreek sprouts, and resulted in implementation of Europe-wide control measures in July 2011.

Published
2012

5. Improved V3 genotyping with duplicate PCR amplification for determining HIV-1 tropism

Author

Hervé Fleury, Bernard Masquelier, P. André, Anne Maillard, Florence Nicot, Stéphanie Raymond, Jacques Izopet, Ali Si-Mohammed, Anne-Geneviève Marcelin, B. Masquelier, Françoise Brun-Vézinet, Michel Segondy, Diane Descamps, Jean-Claude Tardy, Muriel Macé, P. Recordon-Pinson, F. Nicot, V. Calvez, Mary-Anne Trabaud, A G Marcelin, Corinne Amiel, Adrien Saliou, Véronique Schneider, Catherine Tamalet, Pierre Delobel, Brigitte Montes, Mouna Lazrek, Laurence Morand-Joubert, Astrid Vabret, Coralie Pallier, Annick Ruffault, Patricia Recordon-Pinson, C Charpentier, Vincent Calvez, Cathia Soulié, Delphine Desbois, Virginie Ferré, Jacqueline Cottalorda, F. Flandre, S. Raymond, P. Delobel, E. Dussaix, Philippe Flandre, J. Izopet, Centre de Physiopathologie Toulouse Purpan (CPTP), Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université de Toulouse (UT), Laboratoire Virologie [CHU Toulouse], Institut Fédératif de Biologie (IFB), Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Pôle Biologie [CHU Toulouse], Centre Hospitalier Universitaire de Toulouse (CHU Toulouse), Microbiologie Fondamentale et Pathogénicité (MFP), Université Bordeaux Segalen - Bordeaux 2-Centre National de la Recherche Scientifique (CNRS), Service Maladies infectieuses et tropicales [CHU Toulouse], Pôle Inflammation, infection, immunologie et loco-moteur [CHU Toulouse] (Pôle I3LM Toulouse), Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Centre Hospitalier Universitaire de Toulouse (CHU Toulouse), Virology Laboratory, Hospital, Service de microbiologie, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-AP-HP - Hôpital Bichat - Claude Bernard [Paris], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Epidémiologie, stratégies thérapeutiques et virologie cliniques dans l'infection à VIH, Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Unité de Mathématiques Pures et Appliquées (UMPA-ENSL), École normale supérieure de Lyon (ENS de Lyon)-Centre National de la Recherche Scientifique (CNRS), Service de virologie et d'immunologie biologique, CHU Bordeaux [Bordeaux]-Groupe hospitalier Pellegrin, Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université Fédérale Toulouse Midi-Pyrénées, Laboratoire de Virologie, CHU Toulouse [Toulouse]-Hôpital Purpan [Toulouse], CHU Toulouse [Toulouse], Microbiologie cellulaire et moléculaire et pathogénicité (MCMP), Service des Maladies Infectieuses et Tropicales, École normale supérieure - Lyon (ENS Lyon)-Centre National de la Recherche Scientifique (CNRS), and VIROLOGIE

Subjects
Microbiology (medical), Adult, Male, Receptors, CXCR4, Genotype, education, HIV Infections, Biology, MESH: Phenotype, MESH: Receptors, CXCR4, law.invention, MESH: Genotype, MESH: HIV-1, 03 medical and health sciences, 0302 clinical medicine, law, MESH: Reverse Transcriptase Polymerase Chain Reaction, HIV tropism, Humans, Pharmacology (medical), 030212 general & internal medicine, Genotyping, Tropism, Polymerase chain reaction, Pharmacology, 0303 health sciences, MESH: Humans, MESH: Middle Aged, 030306 microbiology, Reverse Transcriptase Polymerase Chain Reaction, MESH: Adult, MESH: HIV Infections, Amplicon, Middle Aged, Virology, Molecular biology, MESH: Male, 3. Good health, Viral Tropism, Infectious Diseases, Phenotype, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Tissue tropism, HIV-1, Female, MESH: Female, MESH: Viral Tropism, Trofile assay
Abstract

International audience; OBJECTIVES: To determine whether genotyping of HIV-1 by duplicate PCR amplification of the region encoding the V3 loop is more sensitive than single PCR for detecting CXCR4-using viruses. PATIENTS AND METHODS: The V3 genotypes of the HIV-1 infecting 152 patients enrolled in the multicentre GenoTropism ANRS study were determined by all the participating laboratories using a single PCR and V3 bulk sequencing. In parallel, one laboratory determined the V3 genotype using duplicate PCR and bulk sequencing of pooled amplicons. HIV tropism was predicted with the geno2pheno10 algorithm. The phenotypes of all samples were determined with the Trofile assay and the Toulouse tropism test (TTT) recombinant virus assay. RESULTS: Geno2pheno10 was 56.8% sensitive and 75.9% specific when compared with the Trofile assay for detecting CXCR4-using viruses after a single PCR. Duplicate amplification and bulk sequencing of the pooled PCR amplicons increased the sensitivity to 68.2% and specificity to 79.6%. Geno2pheno10 was 64.1% sensitive and 77.0% specific when compared with the TTT assay for detecting CXCR4-using viruses after a single PCR. Duplicate amplification and sequencing of the pooled PCR amplicons increased sensitivity to 76.9% and specificity to 80.5%. CONCLUSIONS: The genotypic determination of HIV-1 tropism can be improved by duplicate amplifications and sequencing the pooled PCR products. This is a good compromise between improved sensitivity and reasonable cost for the genotype-based determination of tropism.

Published
2011

6. Outbreak of haemolytic uraemic syndrome and bloody diarrhoea due to Escherichia coli O104:H4, south-west France, June 2011

Author

A Bone, François-Xavier Weill, Jourdan-da Silva N, P. Rolland, Christian Combe, Vendrely B, Patricia Mariani-Kurkdjian, de Valk H, Vaillant, Edouard Bingen, Desjardin M, Delmas Y, d'Andigne E, G. Gault, Muriel Macé, Charron M, L. A. King, Harold Noel, Raymond Bercion, C Castor, B. Aldabe, Nathalie Ong, Cellule interrégionale d’épidémiologie [Nouvelle Aquitaine] (CIRE), Santé publique France - French National Public Health Agency [Saint-Maurice, France], Centre National de Référence - National Reference Center Escherichia coli, Shigella et Salmonella (CNR-ESS), Institut Pasteur [Paris] (IP), AP-HP Hôpital universitaire Robert-Debré [Paris], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Institut de Veille Sanitaire (INVS), European Centre for Disease Prevention and Control [Stockholm, Sweden] (ECDC), CHU Bordeaux [Bordeaux], Hôpital d'Instruction des Armées Robert Picqué, Service de Santé des Armées, Institut Pasteur [Paris], and European Centre for Disease Prevention and Control (ECDC)

Subjects
Adult, Diarrhea, Male, Epidemiology, Bacterial Toxins, Virulence, Microbial Sensitivity Tests, medicine.disease_cause, Polymerase Chain Reaction, Microbiology, Disease Outbreaks, Foodborne Diseases, 03 medical and health sciences, Escherichia coli O104:H4, Antibiotic resistance, Virology, Surveys and Questionnaires, Escherichia coli, Medicine, Humans, Escherichia coli Infections, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, Aged, 0303 health sciences, biology, 030306 microbiology, business.industry, Transmission (medicine), Public Health, Environmental and Occupational Health, Outbreak, Shiga toxin, Drug Resistance, Microbial, Middle Aged, 3. Good health, Immunology, Hemolytic-Uremic Syndrome, biology.protein, Female, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, France, medicine.symptom, business, Gastrointestinal Hemorrhage
Abstract

As of 12:00 28 June 2011, 15 cases of haemolytic uraemic syndrome (HUS) or bloody diarrhoea have been identified in the Gironde, south-west France. Investigations suggest the vehicle of transmission was sprouts, served at an event in Begles on 8 June 2011. A strain of shiga toxin- producing Escherichia coli O104:H4 has been isolated from five cases. This strain is genetically related to the strain identified in the recent E. coli O104:H4 outbreak in Germany, and shares the same virulence and antimicrobial resistance characteristics. .

Published
2011

7. First identification of HIV-1 groups M and O dual infections in Europe

Author

Denys, Brand, Agnès, Beby-Defaux, Muriel, Macé, Sylvie, Brunet, Alain, Moreau, Cendrine, Godet, Xavier, Jais, Françoise, Cazein, Caroline, Semaille, and Francis, Barin

Subjects
Reverse Transcriptase Polymerase Chain Reaction, HIV-1, Humans, RNA, Viral, HIV Infections, France, Serotyping
Published
2004

8. Real-Time PCR as a Versatile Tool for Investigating the Susceptibility of Human Herpesvirus 6 to Antiviral Agents

Author

David Boutolleau, Henri Agut, Stéphanie Précigout, Muriel Macé, Chaysavanh Manichanh, Pascale Bonnafous, and Agnès Gautheret-Dejean

Subjects
Growth kinetics, medicine.drug_class, viruses, Herpesvirus 6, Human, Microbial Sensitivity Tests, Virus Replication, Antiviral Agents, Virus, law.invention, law, medicine, Humans, Pharmacology (medical), Polymerase chain reaction, Cells, Cultured, Pharmacology, Genetics, biology, Reverse Transcriptase Polymerase Chain Reaction, biology.organism_classification, Flow Cytometry, Virology, In vitro, Infectious Diseases, Real-time polymerase chain reaction, Viral replication, DNA, Viral, Human herpesvirus 6, Antiviral drug
Abstract

A quantitative real-time PCR assay was developed for the determination of antiviral drug susceptibility and growth kinetics of human herpesvirus 6. The susceptibility and fitness of a sensitive strain, HST, and its ganciclovir-resistant derivative, GCVR1, were then characterized, leading us to conclude that the mutations of this latter virus did not alter its fitness significantly.

Published
2003

9. No evidence for a major risk of roseolovirus vertical transmission during pregnancy

Author

Agnès Gautheret Dejean, David Boutolleau, Didier Ingrand, Liliane Grangeot Keros, Henri Agut, Denis Cointe, and Muriel Macé

Subjects
Microbiology (medical), Risk, Amniotic fluid, Herpesvirus 6, Human, Roseolovirus Infections, Herpesvirus 7, Human, Polymerase Chain Reaction, law.invention, law, Pregnancy, Medicine, Humans, Dna viral, Pregnancy Complications, Infectious, Polymerase chain reaction, biology, business.industry, biology.organism_classification, medicine.disease, Amniotic Fluid, Virology, Infectious Disease Transmission, Vertical, Infectious Diseases, Transmission (mechanics), DNA, Viral, Female, business, Roseolovirus
Published
2003

10. Therapeutic drug monitoring of HIV protease inhibitors using high-performance liquid chromatography with ultraviolet or photodiode array detection

Author

Jean-Marc Besnier, Gilles Paintaud, Frédéric Bastides, Elisabeth Autret-Leca, Chantal Le Guellec, Patrice Leibenguth, Marie-Laure Gaudet, and Muriel Macé

Subjects
medicine.medical_treatment, Pharmacology, Sensitivity and Specificity, Mixed Function Oxygenases, Amprenavir, Cytochrome P-450 Enzyme System, Indinavir, medicine, HIV Protease Inhibitor, Cytochrome P-450 CYP3A, Humans, Pharmacology (medical), Protease inhibitor (pharmacology), Chromatography, High Pressure Liquid, Chromatography, Protease, medicine.diagnostic_test, Chemistry, HIV Protease Inhibitors, Nelfinavir, Therapeutic drug monitoring, Area Under Curve, Ritonavir, Drug Monitoring, medicine.drug
Abstract

Published data suggest that therapeutic drug monitoring of human immunodeficiency virus protease inhibitors would improve the management of antiretroviral therapy. The authors have developed a high-pressure liquid chromatographic assay allowing simultaneous determination of six protease inhibitors (ritonavir, saquinavir, indinavir, nelfinavir, amprenavir, and lopinavir), using carbamazepine as internal standard. Detection was based on a dual wavelength ultraviolet spectrophotometer and can be improved by the use of a photodiode array detector. Monitoring was performed 1 month after initiation of therapy or in instances of therapeutic failure, side effects, suspicion of noncompliance, drug interactions, or malabsorption. Trough concentrations were 0.15 to 13.6 mg/L for ritonavir, 0.06 to 9.7 mg/L for indinavir, 0.03 to 5.5 mg/L for saquinavir, and 0.15 to 4.15 mg/L for nelfinavir. Concentrations below the limit of quantification were observed in 63/438 (14%) of the patients. Target concentrations are not well established, and reported in vitro inhibitory concentrations may be of limited value. The authors therefore chose to compare observed concentrations with mean plasma concentrations reported in clinical trials. Observed saquinavir and indinavir concentrations were often below or close to these target concentrations, particularly when used as a single protease inhibitor. Concentration-controlled studies should now be used to select proper target concentrations for each protease inhibitor, either prescribed alone or in combination.

Published
2002

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