Determining the analytical specificity of PCR-based assays for the diagnosis of IA: What is Aspergillus?

Détails

ID Serval
serval:BIB_0D858EDADFC8
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Determining the analytical specificity of PCR-based assays for the diagnosis of IA: What is Aspergillus?
Périodique
Medical mycology
Auteur(s)
Morton C.O., White P.L., Barnes R.A., Klingspor L., Cuenca-Estrella M., Lagrou K., Bretagne S., Melchers W., Mengoli C., Caliendo A.M., Cogliati M., Debets-Ossenkopp Y., Gorton R., Hagen F., Halliday C., Hamal P., Harvey-Wood K., Jaton K., Johnson G., Kidd S., Lengerova M., Lass-Florl C., Linton C., Millon L., Morrissey C.O., Paholcsek M., Talento A.F., Ruhnke M., Willinger B., Donnelly J.P., Loeffler J.
Collaborateur(s)
EAPCRI
ISSN
1460-2709 (Electronic)
ISSN-L
1369-3786
Statut éditorial
Publié
Date de publication
01/06/2017
Peer-reviewed
Oui
Volume
55
Numéro
4
Pages
402-413
Langue
anglais
Notes
Publication types: Journal Article ; Multicenter Study
Publication Status: ppublish
Résumé
A wide array of PCR tests has been developed to aid the diagnosis of invasive aspergillosis (IA), providing technical diversity but limiting standardisation and acceptance. Methodological recommendations for testing blood samples using PCR exist, based on achieving optimal assay sensitivity to help exclude IA. Conversely, when testing more invasive samples (BAL, biopsy, CSF) emphasis is placed on confirming disease, so analytical specificity is paramount. This multicenter study examined the analytical specificity of PCR methods for detecting IA by blind testing a panel of DNA extracted from a various fungal species to explore the range of Aspergillus species that could be detected, but also potential cross reactivity with other fungal species. Positivity rates were calculated and regression analysis was performed to determine any associations between technical specifications and performance. The accuracy of Aspergillus genus specific assays was 71.8%, significantly greater (P < .0001) than assays specific for individual Aspergillus species (47.2%). For genus specific assays the most often missed species were A. lentulus (25.0%), A. versicolor (24.1%), A. terreus (16.1%), A. flavus (15.2%), A. niger (13.4%), and A. fumigatus (6.2%). There was a significant positive association between accuracy and using an Aspergillus genus PCR assay targeting the rRNA genes (P = .0011). Conversely, there was a significant association between rRNA PCR targets and false positivity (P = .0032). To conclude current Aspergillus PCR assays are better suited for detecting A. fumigatus, with inferior detection of most other Aspergillus species. The use of an Aspergillus genus specific PCR assay targeting the rRNA genes is preferential.

Mots-clé
Aspergillus/classification, Aspergillus/genetics, Aspergillus/isolation & purification, Humans, Invasive Pulmonary Aspergillosis/diagnosis, Molecular Diagnostic Techniques/methods, Polymerase Chain Reaction/methods, Sensitivity and Specificity, Aspergillus PCR, analytical specificity, cross reactivity, detection range
Pubmed
Web of science
Création de la notice
04/09/2017 18:58
Dernière modification de la notice
20/08/2019 12:34
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