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Evaluation of aspergillus PCR protocols for testing serum specimens

White, P. Lewis, Mengoli, Carlo, Bretagne, Stephane, Cuenca-Estrella, Manual, Finnstrom, Niklas, Klingspor, Lena, Melchers, Willem J. G., McCulloch, Elaine, Barnes, Rosemary Ann, Donnelly, J. Peter and Loeffler, Juergen 2011. Evaluation of aspergillus PCR protocols for testing serum specimens. Journal of Clinical Microbiology 49 (11) , pp. 3842-3848. 10.1128/JCM.05316-11

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Abstract

A panel of human serum spiked with various amounts of A. fumigatus genomic DNA was distributed to 23 centres within the European Aspergillus PCR initiative to determine analytical performance of PCR. Information regarding specific methodological components and PCR performance requested. The information provided was made anonymous and meta-regression analysis 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 was 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 ITS region were shown. Negative associations between sensitivity and the use of larger elution volumes (≥100μl) and PCR targeting the mitochondrial genes was demonstrated. Most Aspergillus PCR protocols used to test serum generate satisfactory analytical performance. Testing serum requires less standardisation and the specific recommendations shown in this manuscript will only improve performance.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Medicine
Subjects: Q Science > QR Microbiology
Additional Information: Pdf uploaded in accordance with publisher's policy at http://www.sherpa.ac.uk/romeo/issn/0095-1137/ (accessed 25/02/2014)
Publisher: American Society for Microbiology
ISSN: 0095-1137
Date of First Compliant Deposit: 30 March 2016
Last Modified: 04 Jun 2017 03:49
URI: http://orca.cf.ac.uk/id/eprint/27438

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