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Article Dans Une Revue Medical Mycology Année : 2021

Interlaboratory evaluation of Mucorales PCR assays for testing serum specimens

1 Service de parasitologie et mycologie [CHRU de Besançon]
2 LCE - Laboratoire Chrono-environnement (UMR 6249)
3 Unipd - Università degli Studi di Padova = University of Padua
4 CNRMA - Centre National de Référence Mycoses Invasives et Antifongiques - National Reference Center Invasive Mycoses & Antifungals
5 Mycologie moléculaire - Molecular Mycology
6 Hôpitaux Universitaire Saint-Louis, Lariboisière, Fernand-Widal
7 UPCité - Université Paris Cité
8 UPEC Médecine - Université Paris-Est Créteil Val-de-Marne - Faculté de médecine
9 ENVA - École nationale vétérinaire d'Alfort
10 CHU Henri Mondor [Créteil]
11 Hôpital Necker - Enfants Malades [AP-HP]
12 BPF - Biologie et Pathogénicité fongiques
13 UNIMI - Università degli Studi di Milano = University of Milan
14 UGSF - Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576
15 Laboratoire de parasitologie mycologie (CHU de Dijon)
16 VAlMiS - Vin Aliment Microbiologie et Stress
17 Laboratoire de parasitologie et de mycologie médicales [CHU Amiens]
18 AGIR - Agents infectieux, résistance et chimiothérapie - UR UPJV 4294
19 Laboratoire de Parasitologie et de Mycologie Médicale [Strasbourg]
20 Leopold Franzens Universität Innsbruck - University of Innsbruck
21 UFR Médecine UPCité - UFR Médecine [Santé] - Université Paris Cité
22 Universiteit Utrecht / Utrecht University [Utrecht]
23 University Medical Center [Utrecht]
24 Shanghai Key Laboratory of Molecular Medical Mycology, Changzheng Hospital
25 Statens Serum Institut [Copenhagen]
26 Service de Parasitologie et Mycologie [CHU Toulouse]
27 CPTP - Centre de Physiopathologie Toulouse Purpan
28 Erasmus MC - Erasmus University Medical Center [Rotterdam]
29 IMU - Innsbruck Medical University = Medizinische Universität Innsbruck
30 University Hospital Brno
31 CHU Strasbourg - Centre Hospitalier Universitaire [Strasbourg]
32 CHU Nantes - Centre hospitalier universitaire de Nantes
33 CHU Clermont-Ferrand
34 University Hospital Wuerzburg / Universitäts­klinikum Würzburg
35 Medizinische Universität Wien = Medical University of Vienna
36 Public Health Wales [Cardiff, Royaume uni]
37 University of Texas Health Science Center at San Antonio [San Antonio, Tx, USA]
C Halliday
  • Fonction : Auteur
C Nourrisson
R Barnes
  • Fonction : Auteur
M Cruciani
  • Fonction : Auteur

Résumé

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.
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hal-03123965 , version 1 (19-01-2024)

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S Rocchi, E Scherer, C Mengoli, A Alanio, F Botterel, et al.. Interlaboratory evaluation of Mucorales PCR assays for testing serum specimens: A study by the fungal PCR Initiative and the Modimucor study group. Medical Mycology, 2021, 59 (2), pp.126-138. ⟨10.1093/mmy/myaa036⟩. ⟨hal-03123965⟩
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