Evaluation of Aspergillus PCR protocols for testing serum specimens.

Details

Serval ID
serval:BIB_48D831699850
Type
Article: article from journal or magazin.
Collection
Publications
Title
Evaluation of Aspergillus PCR protocols for testing serum specimens.
Journal
Journal of clinical microbiology
Author(s)
White P.L., Mengoli C., Bretagne S., Cuenca-Estrella M., Finnstrom N., Klingspor L., Melchers W.J., McCulloch E., Barnes R.A., Donnelly J.P., Loeffler J.
Working group(s)
European Aspergillus PCR Initiative (EAPCRI)
Contributor(s)
Donnelly J.P., Loeffler J., Barnes R.A., Loeffler J., White P.L., Bretagne S., Melchers W., Klingspor L., Finnstrom N., McCulloch E., Williams C., Cuenca-Estrella M., Barnes R.A., Cordonnier C., Maertens J., Klingspor L., Heinz W., Jones B., Mengoli C., Cruciani M., Loeffler J., Barnes R.A., Donnelly J.P., Chrenkova V., Guiver M., Halliday C., Sleiman S., Linton C., Johnson E., Gorton R., Kibbler C., Lengerova M., Mayans E.R., Rogers T., Morton O., Sendid B., Caliendo A., Patterson T., Lagrou K., Wilks M., Moody A., Buitrago M.J., Bernal-Martinez L., Billes J., Hauser P.
ISSN
1098-660X (Electronic)
ISSN-L
0095-1137
Publication state
Published
Issued date
11/2011
Peer-reviewed
Oui
Volume
49
Number
11
Pages
3842-3848
Language
english
Notes
Publication types: Evaluation Studies ; Journal Article
Publication Status: ppublish
Abstract
A panel of human serum samples spiked with various amounts of Aspergillus fumigatus genomic DNA was distributed to 23 centers within the European Aspergillus PCR Initiative to determine analytical performance of PCR. Information regarding specific methodological components and PCR performance was requested. The information provided was made anonymous, and meta-regression analysis was performed to determine any procedural factors that significantly altered PCR performance. Ninety-seven percent of protocols were able to detect a threshold of 10 genomes/ml on at least one occasion, with 83% of protocols reproducibly detecting this concentration. Sensitivity and specificity were 86.1% and 93.6%, respectively. Positive associations between sensitivity and the use of larger sample volumes, an internal control PCR, and PCR targeting the internal transcribed spacer (ITS) region were shown. Negative associations between sensitivity and the use of larger elution volumes (≥100 μl) and PCR targeting the mitochondrial genes were demonstrated. Most Aspergillus PCR protocols used to test serum generate satisfactory analytical performance. Testing serum requires less standardization, and the specific recommendations shown in this article will only improve performance.

Keywords
Aspergillosis/diagnosis, Aspergillosis/microbiology, Aspergillus fumigatus/isolation & purification, Humans, International Cooperation, Molecular Diagnostic Techniques/methods, Molecular Diagnostic Techniques/standards, Mycology/methods, Mycology/standards, Polymerase Chain Reaction/methods, Polymerase Chain Reaction/standards, Reproducibility of Results, Sensitivity and Specificity, Serum/microbiology
Pubmed
Web of science
Open Access
Yes
Create date
11/09/2017 15:43
Last modification date
20/08/2019 13:55
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