title: Development of a real-time RT-PCR assay for a novel influenza A (H1N1) virus.
abstract: 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.
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No. 20 Dongda Street, Fengtai District, Beijing 100071, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Jiang
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Kang
authors.forename: Yongqiang
authors.initials: Y
authors.lastname: Deng
authors.forename: Hui
authors.initials: H
authors.lastname: Zhao
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Li
authors.forename: Xuedong
authors.initials: X
authors.lastname: Yu
authors.forename: Man
authors.initials: M
authors.lastname: Yu
authors.forename: Ede
authors.initials: E
authors.lastname: Qin
authors.forename: Qingyu
authors.initials: Q
authors.lastname: Zhu
authors.forename: Yinhui
authors.initials: Y
authors.lastname: Yang
authors.forename: Chengfeng
authors.initials: C
authors.lastname: Qin
mesh_terms: D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics / Q000302:isolation & purification*; D053122:Influenza A Virus, H3N2 Subtype / Q000235:genetics / Q000302:isolation & purification*; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics / Q000302:isolation & purification*; D007251:Influenza, Human / Q000175:diagnosis* / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity