State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No. 20 Dongda Street, Fengtai District, Beijing 100071, China.
J Virol Methods. 2010 Feb;163(2):470-3. doi: 10.1016/j.jviromet.2009.09.021. Epub 2009 Oct 6.
A pandemic caused by a novel influenza A virus (H1N1) poses a serious public health threat. In this study, a real-time reverse transcriptase PCR (RT-PCR) assay based on the hemagglutinin gene was developed that discriminates the novel H1N1 from swine influenza virus, seasonal H1N1/H3N2 virus and the highly pathogenic H5N1 avian influenza virus. The sensitivity of this assay was 0.2 50% tissue culture infective dose of virus and 200 copies of in vitro-transcribed target RNA. Three hundred and forty-eight clinical specimens from suspected H1N1 patients were tested using this assay, and forty-two (12.07%) were found to be positive. Tests using the real-time PCR assay recommended by WHO and virus isolation gave identical results. This sensitive and specific real-time RT-PCR assay will contribute to the early diagnosis and control of the emerging H1N1 influenza pandemic.
一种由新型甲型 H1N1 流感病毒引起的大流行对公共卫生构成严重威胁。本研究建立了一种基于血凝素基因的实时逆转录聚合酶链反应（RT-PCR）检测方法，可区分新型 H1N1 与猪流感病毒、季节性 H1N1/H3N2 病毒和高致病性 H5N1 禽流感病毒。该方法的灵敏度为 0.250%组织培养感染剂量的病毒和 200 拷贝体外转录的靶 RNA。使用该检测方法对 348 份疑似 H1N1 患者的临床标本进行了检测，发现 42 份（12.07%）呈阳性。使用世界卫生组织推荐的实时 PCR 检测方法和病毒分离的检测结果相同。这种灵敏且特异的实时 RT-PCR 检测方法将有助于对新发的 H1N1 流感大流行进行早期诊断和控制。
