tetano
Editor, Senior Moderator
J Virol Methods. 2014 Jul 24. pii: S0166-0934(14)00290-0. doi: 10.1016/j.jviromet.2014.07.019. [Epub ahead of print]
Development and clinical testing of a simple, low-density gel element array for influenza identification, subtyping, and H275Y detection.
Chandler DP1, Griesemer SB2, Knickerbocker C3, Golova JB3, Lambarqui A3, Perov AN3, Zimmerman C3, Wiles C3, Rudy GB3, St George K2.
Author information
Abstract
The objectives of this study were to develop a user-friendly, gel element microarray test for influenza virus detection, subtyping, and neuraminidase inhibitor resistance detection, assess the performance characteristics of the assay, and perform a clinical evaluation on retrospective nasopharyngeal swab specimens. A streamlined microarray workflow enabled a single user to run up to 24 tests in an 8h shift. The most sensitive components of the test were the primers and probes targeting the A/H1pdm09 HA gene with an analytical limit of detection (LoD) <100 gene copies (gc) per reaction. LoDs for all targets in nasopharyngeal swab samples were ≤1000gc, with the exception of one target in the seasonal A/H1N1 subtype. Seasonal H275Y variants were detectable in a mixed population when present at >5% with wild type virus, while the 2009 pandemic H1N1 H275Y variant was detectable at ≤1% in a mixture with pandemic wild type virus. Influenza typing and subtyping results concurred with those obtained with real-time RT-PCR assays on more than 97% of the samples tested. The results demonstrate that a large panel of single-plex, real-time RT-PCR tests can be translated to an easy-to-use, sensitive, and specific microarray test for potential diagnostic use.
Copyright ? 2014. Published by Elsevier B.V.
KEYWORDS:
Epidemiology; Microarray; Molecular diagnostic; Oseltamivir resistance; Surveillance
PMID:
25066276
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/25066276
Development and clinical testing of a simple, low-density gel element array for influenza identification, subtyping, and H275Y detection.
Chandler DP1, Griesemer SB2, Knickerbocker C3, Golova JB3, Lambarqui A3, Perov AN3, Zimmerman C3, Wiles C3, Rudy GB3, St George K2.
Author information
Abstract
The objectives of this study were to develop a user-friendly, gel element microarray test for influenza virus detection, subtyping, and neuraminidase inhibitor resistance detection, assess the performance characteristics of the assay, and perform a clinical evaluation on retrospective nasopharyngeal swab specimens. A streamlined microarray workflow enabled a single user to run up to 24 tests in an 8h shift. The most sensitive components of the test were the primers and probes targeting the A/H1pdm09 HA gene with an analytical limit of detection (LoD) <100 gene copies (gc) per reaction. LoDs for all targets in nasopharyngeal swab samples were ≤1000gc, with the exception of one target in the seasonal A/H1N1 subtype. Seasonal H275Y variants were detectable in a mixed population when present at >5% with wild type virus, while the 2009 pandemic H1N1 H275Y variant was detectable at ≤1% in a mixture with pandemic wild type virus. Influenza typing and subtyping results concurred with those obtained with real-time RT-PCR assays on more than 97% of the samples tested. The results demonstrate that a large panel of single-plex, real-time RT-PCR tests can be translated to an easy-to-use, sensitive, and specific microarray test for potential diagnostic use.
Copyright ? 2014. Published by Elsevier B.V.
KEYWORDS:
Epidemiology; Microarray; Molecular diagnostic; Oseltamivir resistance; Surveillance
PMID:
25066276
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/25066276