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Duplex real-time RT-PCR assays for the rapid detection and identification of pandemic (H1N1) 2009 and seasonal influenza viruses A/H1, A/H3 and B. (J Clin Microbiol, abstract, RA-1017, edited)
J Clin Microbiol. 2010 Jan 13. [Epub ahead of print]
Duplex real-time RT-PCR assays for the rapid detection and identification of pandemic (H1N1) 2009 and seasonal influenza viruses A/H1, A/H3 and B.
Chidlow G, Harnett G, Williams S, Levy A, Speers D, Smith DW. - Department of Microbiology, Pathwest Laboratory Medicine WA, Nedlands, Australia; School of Biomedical, Biomolecular and Chemical Sciences, University of Western Australia, Nedlands, Australia; School of Pathology and Laboratory Medicine, University of Western Australia, Nedlands, Australia.
Reports of a novel influenza virus type A (H1N1) emerged from the USA and Mexico in April 2009, now designated by the World Health Organization as pandemic (H1N1) 2009. The management of the pandemic in Australia required rapid and reliable testing of large numbers of specimens for the novel influenza strain and differentiation from seasonal influenza strains. A real-time reverse transcriptase PCR (RT-PCR) assay for the detection of pandemic (H1N1) 2009 was designed and used with existing real-time RT-PCR assays for seasonal influenza viruses A and B. MS2 coliphage was added to all samples and amplified as a quality control. Three duplex RT-PCR assays, each containing two primer pairs and corresponding 5' nuclease probes, were initially evaluated on control material and stored samples, and showed high sensitivity and specificity. More than 11000 clinical samples were then tested for influenza A and B matrix gene targets, and specific hemagglutinin gene targets for seasonal influenza A/H1, A/H3 and pandemic A (H1N1) 2009. Minimum sensitivities and specificities were 98.8% and 100% for pandemic (H1N1) 2009, 81.5% and 98.9% for seasonal A/H1, and 96.3% and 99.6% for A/H3. Automated sample extraction facilitated the rapid processing of samples, so that the assays allowed accurate, rapid and cost-effective screening of large numbers of clinical samples.
PMID: 20071557 [PubMed - as supplied by publisher]
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J Clin Microbiol. 2010 Jan 13. [Epub ahead of print]
Duplex real-time RT-PCR assays for the rapid detection and identification of pandemic (H1N1) 2009 and seasonal influenza viruses A/H1, A/H3 and B.
Chidlow G, Harnett G, Williams S, Levy A, Speers D, Smith DW. - Department of Microbiology, Pathwest Laboratory Medicine WA, Nedlands, Australia; School of Biomedical, Biomolecular and Chemical Sciences, University of Western Australia, Nedlands, Australia; School of Pathology and Laboratory Medicine, University of Western Australia, Nedlands, Australia.
Reports of a novel influenza virus type A (H1N1) emerged from the USA and Mexico in April 2009, now designated by the World Health Organization as pandemic (H1N1) 2009. The management of the pandemic in Australia required rapid and reliable testing of large numbers of specimens for the novel influenza strain and differentiation from seasonal influenza strains. A real-time reverse transcriptase PCR (RT-PCR) assay for the detection of pandemic (H1N1) 2009 was designed and used with existing real-time RT-PCR assays for seasonal influenza viruses A and B. MS2 coliphage was added to all samples and amplified as a quality control. Three duplex RT-PCR assays, each containing two primer pairs and corresponding 5' nuclease probes, were initially evaluated on control material and stored samples, and showed high sensitivity and specificity. More than 11000 clinical samples were then tested for influenza A and B matrix gene targets, and specific hemagglutinin gene targets for seasonal influenza A/H1, A/H3 and pandemic A (H1N1) 2009. Minimum sensitivities and specificities were 98.8% and 100% for pandemic (H1N1) 2009, 81.5% and 98.9% for seasonal A/H1, and 96.3% and 99.6% for A/H3. Automated sample extraction facilitated the rapid processing of samples, so that the assays allowed accurate, rapid and cost-effective screening of large numbers of clinical samples.
PMID: 20071557 [PubMed - as supplied by publisher]
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