Influenza A/B virus detection and influenza A virus subtyping with emphasis on the novel H7N9 virus by using multiplex real-time RT-PCR

Rei-Lin Kuo; Shu-Li Yang; Yi-Chun Liu; Lu-Ting Chen; Chee-Keng Mok; Shu-Ming Kuo; Shin-Ru Shih; Kuo-Chien Tsao

doi:10.1016/j.jviromet.2014.07.020

Influenza A/B virus detection and influenza A virus subtyping with emphasis on the novel H7N9 virus by using multiplex real-time RT-PCR

J Virol Methods. 2014 Nov:208:41-6. doi: 10.1016/j.jviromet.2014.07.020. Epub 2014 Jul 24.

Authors

Rei-Lin Kuo¹, Shu-Li Yang², Yi-Chun Liu³, Lu-Ting Chen³, Chee-Keng Mok⁴, Shu-Ming Kuo⁴, Shin-Ru Shih⁵, Kuo-Chien Tsao⁶

Affiliations

¹ Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taiwan; Department of Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Taiwan.
² Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taiwan; Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan, Taiwan.
³ Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan, Taiwan.
⁴ Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taiwan.
⁵ Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taiwan; Department of Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Taiwan; Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan, Taiwan.
⁶ Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taiwan; Department of Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Taiwan; Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan, Taiwan. Electronic address: [email protected].

PMID: 25066277
DOI: 10.1016/j.jviromet.2014.07.020

Abstract

Infections of the novel avian influenza A H7N9 virus cause severe respiratory diseases and death. In this study, to develop highly sensitive methods for differentially detecting the H7N9 virus, multiplex and singular real-time reverse transcription polymerase chain reaction (RT-PCR) assays were established and examined by targeting the H7 and N9 genes of the H7N9 virus. Furthermore, an additional multiplex assay combining previous real time RT-PCR designs was established to subtype the pandemic H1N1, H3, and H5 influenza viruses. Applying the proposed assay system to analyze 100 clinical specimens collected from respiratory infection cases identified influenza A viruses (pandemic H1N1 and H3) in 23 samples. It has been demonstrated that other common respiratory viruses will not be detected by using this platform.

Keywords: Influenza A H7N9 virus; Multiplex real-time reverse transcription polymerase chain reaction (RT-PCR); Rapid and sensitive detection.

Publication types

Evaluation Study
Research Support, Non-U.S. Gov't

MeSH terms

Humans
Influenza A virus / classification
Influenza A virus / genetics
Influenza A virus / isolation & purification*
Influenza B virus / classification
Influenza B virus / genetics
Influenza B virus / isolation & purification*
Influenza, Human / diagnosis*
Influenza, Human / virology
Molecular Diagnostic Techniques / methods*
Multiplex Polymerase Chain Reaction / methods*
Real-Time Polymerase Chain Reaction / methods*
Reverse Transcriptase Polymerase Chain Reaction / methods*
Sensitivity and Specificity