Interlaboratory evaluations of Mucorales qPCR assays were developed to assess the reproducibility and performance of methods currently used. The participants comprised 12 laboratories from French university hospitals (nine of them participating in the Modimucor study) and 11 laboratories participating in the Fungal PCR Initiative. For panel 1, three sera were each spiked with DNA from three different species (Rhizomucor pusillus, Lichtheimia corymbifera, Rhizopus oryzae). For panel 2, six sera with three concentrations of R. pusillus and L. corymbifera (1, 10, and 100 genomes/ml) were prepared. Each panel included a blind negative-control serum. A form was distributed with each panel to collect results and required technical information, including DNA extraction method, sample volume used, DNA elution volume, qPCR method, qPCR template input volume, qPCR total reaction volume, qPCR platform, and qPCR reagents used. For panel 1, assessing 18 different protocols, qualitative results (positive or negative) were correct in 97% of cases (70/72). A very low interlaboratory variability in Cq values (SD = 1.89 cycles) were observed. For panel 2 assessing 26 different protocols, the detection rates were high (77-100%) for 5/6 of spiked serum. There was a significant association between the qPCR platform and performance. However, certain technical steps and optimal combinations of factors may also impact performance. The good reproducibility and performance demonstrated in this study support the use of Mucorales qPCR as part of the diagnostic strategy for mucormycosis.

Centre de Physiopathologie de Toulouse Purpan Université de Toulouse CNRS INSERM UPS Toulouse France

Clinical Mycology Reference Laboratory Centre for Infectious Diseases and Microbiology Laboratory Services ICPMR NSW Health Pathology Westmead NSW 2145 Australia

Département de Parasitologie et Mycologie Médicale EA1155 IICiMed Nantes Université Nantes France

Department of Internal Medicine 2 WÜ4i University Hospital Wuerzburg Wuerzburg Germany

Department of Internal Medicine Hematology and Oncology University Hospital Brno Brno Czech Republic

Department of Medical Microbiology and Infectious Diseases Erasmus MC University Medical Center Rotterdam The Netherlands

Department of Medical Microbiology University Medical Center Utrecht Utrecht The Netherlands

Division of Clinical Microbiology Department of Laboratory Medicine Medical University of Vienna

Division of Infectious Diseases San Antonio Center for Medical Mycology San Antonio Texas United States of America

EA Dynamyc 7380 UPEC ENVA Faculté de Médecine de Créteil 8 rue du Général Sarrail 94010 Créteil France

Equipe AGIR Agents Infectieux Résistance et Chimiothérapie UR4294 Université de Picardie Jules Verne Amiens France

Fungal Biology and Pathogenicity Unit INRA USC 2019 Institut Pasteur Paris France

Infectious Diseases Unit ULSS 20 Verona Italy

Inserm U1285 Univ Lille UMR CNRS 8576 UGSF Unité de Glycobiologie Structurale et Fonctionnelle F 59000 Lille France

Institut for Hygiene and Medical Microbiology Medical University of Innsbruck Austria

Institut Pasteur CNRS National Reference Center for Invasive Mycoses and Antifungals Molecular Mycology Unit UMR2000 Paris France

Institute of Hygiene and Medical Microbiology Medical University of Innsbruck Innsbruck Austria

Lab Medical Mycology Dip Scienze Biomediche per la Salute Università degli Studi di Milano Milano Italy

Laboratoire de Parasitologie et de Mycologie Médicale Hôpitaux Universitaires de Strasbourg 1 Place de l'Hôpital 67000 Strasbourg France

Laboratoire de Parasitologie et Mycologie Médicales Centre de Biologie Humaine CHU Amiens Picardie France

Laboratoire de Parasitologie Mycologie CHU Clermont Ferrand 3IHP France

Laboratoire de Parasitologie Mycologie CHU Nantes Nantes France

Laboratoire de Parasitologie Mycologie Plateforme de Biologie Hospitalo Universitaire Gérard Mack Dijon France

Laboratory of Medical Mycology Jining No 1 People's Hospital Jining Shandong People's Republic of China

Medical Microbiology and Infectious Diseases Cardiff University School of Medicine Cardiff United Kingdom

Molecular Medicine University of Padova Padova Italy

Mycology Reference Laboratory Public Health Wales Microbiology Cardiff United Kingdom

Mycology Unit Department for Bacteria Parasites and Fungi Statens Serum Institut Copenhagen Denmark

Parasitology Mycology Laboratory Lariboisière Saint Louis Fernand Widal hospitals Assistance Publique Hôpitaux de Paris Paris France

Parasitology Mycology Unit Necker Enfants Malades Hospital APHP Paris France

Parasitology Mycology University Hospital Besançon Besançon France

Service de Parasitologie Mycologie CHU Toulouse Toulouse France

UMR PAM Univ Bourgogne Franche Comté AgroSup Dijon Equipe Vin Aliment Microbiologie Stress Dijon France

UMR6249 CNRS Chrono Environnement University of Bourgogne Franche Comté Besançon Besançon France

Unité de Parasitologie Mycologie Département de Virologie Bactériologie Hygiène Mycologie Parasitologie DHU VIC CHU Henri Mondor AP HP 51 avenue du Maréchal de Lattre de Tassigny 94010 Créteil France

Université de Paris France

Westerdijk Fungal Biodiversity Institute Uppsalalaan 8 3584 CT Utrecht The Netherlands

Citace poskytuje Crossref.org

Interlaboratory assays from the fungal PCR Initiative and the Modimucor Study Group to improve qPCR detection of Mucorales DNA in serum: one more step toward standardization