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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.
AuthorsYan Li, Tao Wu, Xian Qi, Yiyue Ge, Xiling Guo, Bin Wu, Huiyan Yu, Yefei Zhu, Zhiyang Shi, Hua Wang, Lunbiao Cui, Minghao Zhou
JournalJournal of virological methods (J Virol Methods) Vol. 194 Issue 1-2 Pg. 194-6 (Dec 2013) ISSN: 1879-0984 [Electronic] Netherlands
PMID24001700 (Publication Type: Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't)
CopyrightCopyright © 2013 Elsevier B.V. All rights reserved.
Chemical References
  • Hemagglutinin Glycoproteins, Influenza Virus
  • Viral Proteins
  • NA protein, influenza A virus
  • Neuraminidase
Topics
  • China
  • Environmental Microbiology
  • Hemagglutinin Glycoproteins, Influenza Virus (genetics)
  • Humans
  • Influenza A Virus, H7N9 Subtype (genetics, isolation & purification)
  • Influenza, Human (diagnosis, virology)
  • Mass Screening (methods)
  • Multiplex Polymerase Chain Reaction (methods)
  • Neuraminidase (genetics)
  • Real-Time Polymerase Chain Reaction (methods)
  • Reverse Transcriptase Polymerase Chain Reaction (methods)
  • Sensitivity and Specificity
  • Time Factors
  • Viral Proteins (genetics)
  • Virology (methods)

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