Giuseppe
Emeritus
J Clin Microbiol. 2009 Sep 30. [Epub ahead of print]
Switching gears for an influenza pandemic: validation of a duplex RT-PCR for simultaneous detection and confirmation of pandemic (H1N1) 2009.
Leblanc JJ, Li Y, Bastien N, Forward KR, Davidson RJ, Hatchette TF. - Department of Pathology and Laboratory Medicine, Queen Elizabeth II Health Science Center, Halifax, Nova Scotia, Canada; Department of Pathology, Dalhousie University, Halifax, Nova Scotia, Canada; National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada.
Rapid methods for detection and confirmation of pandemic influenza A [also known as pandemic (H1N1) 2009] are of utmost importance. In this study, a conventional reverse-transcriptase polymerase chain reaction (RT-PCR) was designed, optimized and validated for the detection of influenza A and the hemagglutinin of swine lineage H1 (swH1). Nucleic acids were extracted from 198 consecutive nasopharyngeal, nasal or throat swabs collected early in the outbreak [127 negatives, 66 pandemic (H1N1) 2009, 3 seasonal influenza A (H1N1) and 2 seasonal influenza A (H3N2)]. The performance characteristics of duplex RT-PCR were assessed compared to various detection methods: monoplex RT-PCR at the National Microbiology Laboratory (NML), a real-time RT-PCR using a Center for Disease Control and Prevention (CDC) protocol, a in-house multiplex RT-PCR [targeting influenza A, influenza B and respiratory syncytial virus (RSV)], and a rapid antigen test, the BinaxNOW Influenza A & B. For influenza A detection, the sensitivity of duplex RT-PCR was 97.2%, versus 74.6%, 71.8%, 47.8%, and 12.7% for the other assays, respectively. Duplex RT-PCR was also able to identify swH1 in the 94% of cases, thereby reducing the number of specimens forwarded to reference laboratories for confirmation. Only a limited number of influenza A-positive specimens fell below the limit of detection of the swH1 primers. Overall, duplex RT-PCR is a reliable method for simultaneous detection and confirmation of pandemic (H1N1) 2009 and would be particularly attractive to laboratories without real-time RT-PCR capability.
PMID: 19794033 [PubMed - as supplied by publisher]
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Switching gears for an influenza pandemic: validation of a duplex RT-PCR for simultaneous detection and confirmation of pandemic (H1N1) 2009.
Leblanc JJ, Li Y, Bastien N, Forward KR, Davidson RJ, Hatchette TF. - Department of Pathology and Laboratory Medicine, Queen Elizabeth II Health Science Center, Halifax, Nova Scotia, Canada; Department of Pathology, Dalhousie University, Halifax, Nova Scotia, Canada; National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada.
Rapid methods for detection and confirmation of pandemic influenza A [also known as pandemic (H1N1) 2009] are of utmost importance. In this study, a conventional reverse-transcriptase polymerase chain reaction (RT-PCR) was designed, optimized and validated for the detection of influenza A and the hemagglutinin of swine lineage H1 (swH1). Nucleic acids were extracted from 198 consecutive nasopharyngeal, nasal or throat swabs collected early in the outbreak [127 negatives, 66 pandemic (H1N1) 2009, 3 seasonal influenza A (H1N1) and 2 seasonal influenza A (H3N2)]. The performance characteristics of duplex RT-PCR were assessed compared to various detection methods: monoplex RT-PCR at the National Microbiology Laboratory (NML), a real-time RT-PCR using a Center for Disease Control and Prevention (CDC) protocol, a in-house multiplex RT-PCR [targeting influenza A, influenza B and respiratory syncytial virus (RSV)], and a rapid antigen test, the BinaxNOW Influenza A & B. For influenza A detection, the sensitivity of duplex RT-PCR was 97.2%, versus 74.6%, 71.8%, 47.8%, and 12.7% for the other assays, respectively. Duplex RT-PCR was also able to identify swH1 in the 94% of cases, thereby reducing the number of specimens forwarded to reference laboratories for confirmation. Only a limited number of influenza A-positive specimens fell below the limit of detection of the swH1 primers. Overall, duplex RT-PCR is a reliable method for simultaneous detection and confirmation of pandemic (H1N1) 2009 and would be particularly attractive to laboratories without real-time RT-PCR capability.
PMID: 19794033 [PubMed - as supplied by publisher]
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