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Interlaboratory evaluation of Mucorales PCR assays for testing serum specimens: A study by the fungal PCR Initiative and the Modimucor study group
S. Rocchi, E. Scherer, C. Mengoli, A. Alanio, F. Botterel, ME. Bougnoux, S. Bretagne, M. Cogliati, M. Cornu, F. Dalle, C. Damiani, J. Denis, S. Fuchs, M. Gits-Muselli, F. Hagen, C. Halliday, R. Hare, X. Iriart, C. Klaassen, M. Lackner, M....
Jazyk angličtina Země Velká Británie
Typ dokumentu hodnotící studie, časopisecké články, multicentrická studie
PubMed
32534456
DOI
10.1093/mmy/myaa036
Knihovny.cz E-zdroje
- MeSH
- diagnostické techniky molekulární normy MeSH
- DNA fungální genetika MeSH
- klinické laboratorní techniky přístrojové vybavení metody normy MeSH
- kvantitativní polymerázová řetězová reakce normy MeSH
- lidé MeSH
- Mucorales genetika MeSH
- mukormykóza krev diagnóza MeSH
- nemocnice univerzitní statistika a číselné údaje MeSH
- odchylka pozorovatele MeSH
- reprodukovatelnost výsledků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- multicentrická studie MeSH
- Geografické názvy
- Francie MeSH
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
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 University Medical Center Utrecht Utrecht The Netherlands
Division of Clinical Microbiology Department of Laboratory Medicine Medical University of Vienna
Fungal Biology and Pathogenicity Unit INRA USC 2019 Institut Pasteur Paris France
Infectious Diseases Unit ULSS 20 Verona Italy
Institut for Hygiene and Medical Microbiology Medical University of Innsbruck Austria
Institute of Hygiene and Medical Microbiology Medical University of Innsbruck Innsbruck Austria
Laboratoire de Parasitologie Mycologie CHU Clermont Ferrand 3IHP France
Laboratoire de Parasitologie Mycologie CHU Nantes Nantes France
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 Unit Necker Enfants Malades Hospital APHP Paris France
Parasitology Mycology University Hospital Besançon Besançon France
Service de Parasitologie Mycologie CHU Toulouse Toulouse France
UMR6249 CNRS Chrono Environnement University of Bourgogne Franche Comté Besançon Besançon France
Westerdijk Fungal Biodiversity Institute Uppsalalaan 8 3584 CT Utrecht The Netherlands
Citace poskytuje Crossref.org
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