title: Simultaneous detection of hemagglutinin and neuraminidase genes of novel influenza A (H7N9) by duplex real-time reverse transcription polymerase chain reaction.
abstract: A novel reassortant influenza A (H7N9) virus emerged recently in China. In this study, a duplex real-time reverse transcription polymerase chain reaction (rRT-PCR) assay was developed for the simultaneous detection of hemagglutinin (HA) and neuraminidase (NA) genes of H7N9 influenza viruses. The sensitivity of the assay was determined to be 10 RNA copies per reaction for both HA and NA genes. No cross-reactivity was observed with other influenza virus subtypes or respiratory tract viruses. One hundred and forty-six clinical and environmental specimens were tested and compared with reference methods and were found to be consistent. The assay is suitable for large-scale screening due to short turnaround times and high specificity, sensitivity, and reproducibility.
authors.affiliation: Key Laboratory of Enteric Pathogenic Microbiology, Ministry of Health, Institute of Pathogenic Microbiology, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing 210009, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Tao
authors.initials: T
authors.lastname: Wu
authors.forename: Xian
authors.initials: X
authors.lastname: Qi
authors.forename: Yiyue
authors.initials: Y
authors.lastname: Ge
authors.forename: Xiling
authors.initials: X
authors.lastname: Guo
authors.forename: Bin
authors.initials: B
authors.lastname: Wu
authors.forename: Huiyan
authors.initials: H
authors.lastname: Yu
authors.forename: Yefei
authors.initials: Y
authors.lastname: Zhu
authors.forename: Zhiyang
authors.initials: Z
authors.lastname: Shi
authors.forename: Hua
authors.initials: H
authors.lastname: Wang
authors.forename: Lunbiao
authors.initials: L
authors.lastname: Cui
authors.forename: Minghao
authors.initials: M
authors.lastname: Zhou
mesh_terms: D002681:China; D004783:Environmental Microbiology; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics*; D006801:Humans; D064766:Influenza A Virus, H7N9 Subtype / Q000235:genetics / Q000302:isolation & purification*; D007251:Influenza, Human / Q000175:diagnosis* / Q000821:virology; D008403:Mass Screening / Q000379:methods; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D009439:Neuraminidase / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors; D014764:Viral Proteins / Q000235:genetics*; D014773:Virology / Q000379:methods
chemical_list: D019267:Hemagglutinin Glycoproteins, Influenza Virus; D014764:Viral Proteins; C487630:NA protein, influenza A virus; D009439:Neuraminidase
keywords: H7N9; Influenza A; Real-time reverse transcription polymerase chain reaction