Abstract
BackgroundInfluenza A, including avian influenza, is a major public health threat in developed and developing countries. Rapid and accurate detection is a key component of strategies to contain spread of infection, and the efficient diagnosis of influenza-like-illness is essential to protect health infrastructure in the event of a major influenza outbreak.MethodsWe developed a multiplexed PCR (MT-PCR) assay for the simultaneous diagnosis of respiratory viruses causing influenza-like illness, including the specific recognition of influenza A haemagglutinin subtypes H1, H3, and H5. We tested several hundred clinical specimens in two diagnostic reference laboratories and compared the results with standard techniques.ResultsThe sensitivity and specificity of these assays was higher than individual assays based on direct antigen detection and standard PCR against a range of control templates and in several hundred clinical specimens. The MT-PCR assays provided differential diagnoses as well as potentially useful quantitation of virus in clinical samples.ConclusionsMT-PCR is a potentially powerful tool for the differential diagnosis of influenza-like illness in the clinical diagnostic laboratory.
Highlights
Influenza A, including avian influenza, is a major public health threat in developed and developing countries
Approximately one billion cases of influenza occur around the world every year, despite the availability of effective vaccines
The initial H5 target is a consensus for clades 1-3 of the highly pathogenic H5N1 strains, consistent with WHO guidelines http://www.who.int/csr/disease/avian_influenza/guidelines/RecAIlabtestsAug07.pdf, but the region targeted by our consensus INF-A primers varies significantly from that of the highly pathogenic avian strains of Influenza A, and this was reflected in a relatively low sensitivity in the INF-A assay for H5N1
Summary
We developed a multiplexed PCR (MT-PCR) assay for the simultaneous diagnosis of respiratory viruses causing influenza-like illness, including the specific recognition of influenza A haemagglutinin subtypes H1, H3, and H5. We tested several hundred clinical specimens in two diagnostic reference laboratories and compared the results with standard techniques
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