Title :

The Fungal PCR Initiative’s evaluation of in-house and commercial Pneumocystis jirovecii qPCR assays: towards a standard for a diagnostics assay

Author(s) :

Gits-Muselli, Maud [Auteur]
Laboratoire de Parasitologie-Mycologie [CHU Saint Louis, Paris]
Mycologie moléculaire - Molecular Mycology
White, P. Lewis [Auteur]
International University of Health and Welfare Hospital [IUHW Hospital]
Mengoli, Carlo [Auteur]
Università degli Studi di Padova = University of Padua [Unipd]
Chen, Sharon [Auteur]
The University of Sydney
Crowley, Brendan [Auteur]
St James's University Hospital
Dingemans, Gijs [Auteur]
Fréalle, Emilie [Auteur]
Centre d’Infection et d’Immunité de Lille - INSERM U 1019 - UMR 9017 - UMR 8204 [CIIL]
Centre Hospitalier Régional Universitaire [CHU Lille] [CHRU Lille]
Gorton, Rebecca [Auteur]
Guiver, Malcom [Auteur]
Manchester University NHS Foundation Trust [MFT]
Hagen, Ferry [Auteur]
University Medical Center [Utrecht] [UMCU]
Halliday, Catriona [Auteur]
The University of Sydney
Johnson, Gemma [Auteur]
Lagrou, Katrien [Auteur]
Universitair Ziekenhuis Leuven = University Hospital of Leuven = Hopital universitaire de Louvain [UZ Leuven]
Lengerova, Martina [Auteur]
Mendel University in Brno [MENDELU]
Melchers, Willem Jg [Auteur]
Radboud University Medical Center [Nijmegen] [RadboudUMC]
Novak-Frazer, Lily [Auteur]
Manchester University NHS Foundation Trust [MFT]
Rautemaa-Richardson, Riina [Auteur]
Manchester University NHS Foundation Trust [MFT]
Scherer, Emeline [Auteur]
Centre Hospitalier Régional Universitaire de Besançon [CHRU Besançon]
Steinmann, Joerg [Auteur]
Universität Duisburg-Essen = University of Duisburg-Essen [Essen]
Cruciani, Mario [Auteur]
Ospedale Fracastoro San Bonifacio [Verona]
Barnes, Rosemary [Auteur]
Cardiff University
Donnelly, J. Peter [Auteur]
Radboud University [Nijmegen]
Loeffler, Juergen [Auteur]
Julius-Maximilians-Universität Würzburg = University of Würzburg [Würsburg, Germany] [JMU]
Bretagne, Stéphane [Auteur]
Laboratoire de Parasitologie-Mycologie [CHU Saint Louis, Paris]
Mycologie moléculaire - Molecular Mycology
Alanio, Alexandre [Auteur]
Laboratoire de Parasitologie-Mycologie [CHU Saint Louis, Paris]
Mycologie moléculaire - Molecular Mycology

HAL domain(s) :

Sciences du Vivant [q-bio]/Microbiologie et Parasitologie/Mycologie
Sciences du Vivant [q-bio]/Biodiversité/Systématique, phylogénie et taxonomie
Sciences du Vivant [q-bio]/Médecine humaine et pathologie/Maladies infectieuses

English abstract : [en]

Quantitative real-time PCR (qPCR) is increasingly used to detect Pneumocystis jirovecii for the diagnosis of Pneumocystis pneumonia (PCP), but there are differences in the nucleic acids targeted, DNA only ...
Show more >Quantitative real-time PCR (qPCR) is increasingly used to detect Pneumocystis jirovecii for the diagnosis of Pneumocystis pneumonia (PCP), but there are differences in the nucleic acids targeted, DNA only versus whole nucleic acid (WNA), and also the target genes for amplification. Through the Fungal qPCR Initiative, a working group of the International Society for Human and Animal Mycology, a multicentre and monocentre evaluation of PCP qPCR assays was performed. For the multicentre study, 16 reference laboratories from eight different countries, performing 20 assays analysed a panel consisting of two negative and three PCP positive samples. Aliquots were prepared by pooling residual material from 20 negative or positive- P. jirovecii bronchoalveolar lavage fluids (BALFs). The positive pool was diluted to obtain three concentrations (pure 1:1; 1:100; and 1:1000 to mimic high, medium, and low fungal loads respectively). The monocentre study compared five in-house and five commercial qPCR assays testing 19 individual BALFs on the same amplification platform. Across both evaluations and for all fungal loads, targeting WNA and the mitochondrial small sub-unit (mtSSU) provided the earliest Cq values, compared to only targeting DNA and the mitochondrial large subunit, the major surface glycoprotein or the beta-tubulin genes. Thus, Reverse Transcriptase-qPCR targeting the mtSSU gene could serve as a basis for standardizing the P. jirovecii load, which is essential if qPCR is to be incorporated into clinical care pathways as the reference method, accepting that additional parameters such as amplification platforms still need evaluationShow less >

Language :

Anglais

Collections :

• Centre d'Infection et d'Immunité de Lille (CIIL) - U1019 - UMR 9017

Source :

Harvested from HAL