tetano
Editor, Senior Moderator
Mol Cell Probes. 2011 Oct 17. [Epub ahead of print]
Economic high-throughput-identification of influenza A subtypes from clinical specimens with a DNA-oligonucleotide microarray in an outbreak situation.
Bandt D, Monecke S, Scott C, Gall A, Hoffmann B, Ehricht R.
Source
Institute for Medical Diagnostics Oderland, Frankfurt (Oder), Germany.
Abstract
Influenza A surface proteins H (haemagglutinin) and N (neuraminidase) occur in 16 and 9 distinct genotypes, respectively. The need for a timely production of vaccinations in case of pandemics or seasonal epidemics requires rapid typing methods for the determination of these alleles. The aim of the present study was to develop and improve a rapid and economic assay for determining H and N subtypes of influenza A from patient samples. The assay is based on the hybridisation of labelled amplicons from H and N reverse transcriptase-PCRs using consensus primer pairs to subtype-specific probes on microtiterstripe-mounted DNA-microarrays. An algorithm for semi-automatic data interpretation of raw data and assignment to H and N subtypes was proposed. Altogether, 191 samples were genotyped. This included 134 patient and 44 reference samples as well as controls. Under routine conditions sensitivity and specificity proved to be comparable to conventional nested or real-time PCRs. At least 130 out of 147 array-positive samples were unambiguously assignable. This included all sixteen variants of H as well as all nine variants of N. Furthermore, eighty-two samples from the 2009/2010 "novel H1N1/swine flu" (SF)-outbreak were correctly identified.
Copyright ? 2011. Published by Elsevier Ltd.
PMID:
22019422
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/22019422
Economic high-throughput-identification of influenza A subtypes from clinical specimens with a DNA-oligonucleotide microarray in an outbreak situation.
Bandt D, Monecke S, Scott C, Gall A, Hoffmann B, Ehricht R.
Source
Institute for Medical Diagnostics Oderland, Frankfurt (Oder), Germany.
Abstract
Influenza A surface proteins H (haemagglutinin) and N (neuraminidase) occur in 16 and 9 distinct genotypes, respectively. The need for a timely production of vaccinations in case of pandemics or seasonal epidemics requires rapid typing methods for the determination of these alleles. The aim of the present study was to develop and improve a rapid and economic assay for determining H and N subtypes of influenza A from patient samples. The assay is based on the hybridisation of labelled amplicons from H and N reverse transcriptase-PCRs using consensus primer pairs to subtype-specific probes on microtiterstripe-mounted DNA-microarrays. An algorithm for semi-automatic data interpretation of raw data and assignment to H and N subtypes was proposed. Altogether, 191 samples were genotyped. This included 134 patient and 44 reference samples as well as controls. Under routine conditions sensitivity and specificity proved to be comparable to conventional nested or real-time PCRs. At least 130 out of 147 array-positive samples were unambiguously assignable. This included all sixteen variants of H as well as all nine variants of N. Furthermore, eighty-two samples from the 2009/2010 "novel H1N1/swine flu" (SF)-outbreak were correctly identified.
Copyright ? 2011. Published by Elsevier Ltd.
PMID:
22019422
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/22019422