tetano
Editor, Senior Moderator
J Mol Diagn. 2010 Aug 5. [Epub ahead of print]
Diagnostic Testing for Pandemic Influenza in Singapore. A Novel Dual-Gene Quantitative Real-Time RT-PCR for the Detection of Influenza A/H1N1/2009.
Lee HK, Lee CK, Loh TP, Tang JW, Chiu L, Tambyah PA, Sethi SK, Koay ES.
From the Molecular Diagnosis Centre,* Clinical Chemistry, Microbiology, the Department of Laboratory Medicine, National University Hospital, Singapore; and the Departments of Medicine, and Pathology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
Abstract
With the relative global lack of immunity to the pandemic influenza A/H1N1/2009 virus that emerged in April 2009 as well as the sustained susceptibility to infection, rapid and accurate diagnostic assays are essential to detect this novel influenza A variant. Among the molecular diagnostic methods that have been developed to date, most are in tandem monoplex assays targeting either different regions of a single viral gene segment or different viral gene segments. We describe a dual-gene (duplex) quantitative real-time RT-PCR method selectively targeting pandemic influenza A/H1N1/2009. The assay design includes a primer-probe set specific to only the hemagglutinin (HA) gene of this novel influenza A variant and a second set capable of detecting the nucleoprotein (NP) gene of all swine-origin influenza A virus. In silico analysis of the specific HA oligonucleotide sequence used in the assay showed that it targeted only the swine-origin pandemic strain; there was also no cross-reactivity against a wide spectrum of noninfluenza respiratory viruses. The assay has a diagnostic sensitivity and specificity of 97.7% and 100%, respectively, a lower detection limit of 50 viral gene copies/PCR, and can be adapted to either a qualitative or quantitative mode. It was first applied to 3512 patients with influenza-like illnesses at a tertiary hospital in Singapore, during the containment phase of the pandemic (May to July 2009).
PMID: 20688908 [PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/20688908
Diagnostic Testing for Pandemic Influenza in Singapore. A Novel Dual-Gene Quantitative Real-Time RT-PCR for the Detection of Influenza A/H1N1/2009.
Lee HK, Lee CK, Loh TP, Tang JW, Chiu L, Tambyah PA, Sethi SK, Koay ES.
From the Molecular Diagnosis Centre,* Clinical Chemistry, Microbiology, the Department of Laboratory Medicine, National University Hospital, Singapore; and the Departments of Medicine, and Pathology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
Abstract
With the relative global lack of immunity to the pandemic influenza A/H1N1/2009 virus that emerged in April 2009 as well as the sustained susceptibility to infection, rapid and accurate diagnostic assays are essential to detect this novel influenza A variant. Among the molecular diagnostic methods that have been developed to date, most are in tandem monoplex assays targeting either different regions of a single viral gene segment or different viral gene segments. We describe a dual-gene (duplex) quantitative real-time RT-PCR method selectively targeting pandemic influenza A/H1N1/2009. The assay design includes a primer-probe set specific to only the hemagglutinin (HA) gene of this novel influenza A variant and a second set capable of detecting the nucleoprotein (NP) gene of all swine-origin influenza A virus. In silico analysis of the specific HA oligonucleotide sequence used in the assay showed that it targeted only the swine-origin pandemic strain; there was also no cross-reactivity against a wide spectrum of noninfluenza respiratory viruses. The assay has a diagnostic sensitivity and specificity of 97.7% and 100%, respectively, a lower detection limit of 50 viral gene copies/PCR, and can be adapted to either a qualitative or quantitative mode. It was first applied to 3512 patients with influenza-like illnesses at a tertiary hospital in Singapore, during the containment phase of the pandemic (May to July 2009).
PMID: 20688908 [PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/20688908