Molecular Detection of a Novel Human Influenza (H1N1) of Pandemic Potential by Conventional and Real-Time

BACKGROUND: Influenza A viruses aremedically impor-tant viral pathogens that cause significant mortality andmorbidity throughout theworld. The recent emer-gence of a novel human influenza A virus (H1N1) poses a serious health threat. Molecular tests for rapid detection of this virus are urgently needed. METHODS: We developed a conventional 1-step RT-PCR assay and a 1-step quantitative real-timeRT-PCRassay to detect the novel H1N1 virus, but not the seasonal H1N1 viruses. We also developed an additional real-time RT-PCR that can discriminate the novel H1N1 from other swine and humanH1 subtype viruses. RESULTS: All of the assays had detection limits for the pos-itive control in the range of 1.0 104 to 2.0 103 of themedian tissueculture infectivedose.Assay specificitie