Multiplex reverse transcription-PCR for simultaneous surveillance of influenza A and B viruses

Bin Zhou, Yi Mo Deng, John R. Barnes, October M. Sessions, Tsui Wen Chou, Malania Wilson, Thomas J. Stark, Michelle Volk, Natalie Spirason, Rebecca A. Halpin, Uma Sangumathi Kamaraj, Tao Ding, Timothy B. Stockwell, Mirella Salvatore, Elodie Ghedin, Ian G. Barr, David E. Wentworth

Research output: Contribution to journalArticle

Abstract

Influenza A and B viruses are the causative agents of annual influenza epidemics that can be severe, and influenza A viruses intermittently cause pandemics. Sequence information from influenza virus genomes is instrumental in determining mechanisms underpinning antigenic evolution and antiviral resistance. However, due to sequence diversity and the dynamics of influenza virus evolution, rapid and high-throughput sequencing of influenza viruses remains a challenge. We developed a single-reaction influenza A/B virus (FluA/B) multiplex reverse transcription-PCR (RTPCR) method that amplifies the most critical genomic segments (hemagglutinin [HA], neuraminidase [NA], and matrix [M]) of seasonal influenza A and B viruses for next-generation sequencing, regardless of viral type, subtype, or lineage. Herein, we demonstrate that the strategy is highly sensitive and robust. The strategy was validated on thousands of seasonal influenza A and B virus-positive specimens using multiple next-generation sequencing platforms.

Original languageEnglish (US)
Pages (from-to)3492-3501
Number of pages10
JournalJournal of Clinical Microbiology
Volume55
Issue number12
DOIs
StatePublished - Dec 1 2017

Fingerprint

Influenza B virus
Influenza A virus
Reverse Transcription
Polymerase Chain Reaction
Orthomyxoviridae
Hemagglutinins
Neuraminidase
Pandemics
Human Influenza
Antiviral Agents
Genome

Keywords

  • Influenza
  • NGS
  • RT-PCR
  • Surveillance

ASJC Scopus subject areas

  • Microbiology (medical)

Cite this

Zhou, B., Deng, Y. M., Barnes, J. R., Sessions, O. M., Chou, T. W., Wilson, M., ... Wentworth, D. E. (2017). Multiplex reverse transcription-PCR for simultaneous surveillance of influenza A and B viruses. Journal of Clinical Microbiology, 55(12), 3492-3501. https://doi.org/10.1128/JCM.00957-17

Multiplex reverse transcription-PCR for simultaneous surveillance of influenza A and B viruses. / Zhou, Bin; Deng, Yi Mo; Barnes, John R.; Sessions, October M.; Chou, Tsui Wen; Wilson, Malania; Stark, Thomas J.; Volk, Michelle; Spirason, Natalie; Halpin, Rebecca A.; Kamaraj, Uma Sangumathi; Ding, Tao; Stockwell, Timothy B.; Salvatore, Mirella; Ghedin, Elodie; Barr, Ian G.; Wentworth, David E.

In: Journal of Clinical Microbiology, Vol. 55, No. 12, 01.12.2017, p. 3492-3501.

Research output: Contribution to journalArticle

Zhou, B, Deng, YM, Barnes, JR, Sessions, OM, Chou, TW, Wilson, M, Stark, TJ, Volk, M, Spirason, N, Halpin, RA, Kamaraj, US, Ding, T, Stockwell, TB, Salvatore, M, Ghedin, E, Barr, IG & Wentworth, DE 2017, 'Multiplex reverse transcription-PCR for simultaneous surveillance of influenza A and B viruses', Journal of Clinical Microbiology, vol. 55, no. 12, pp. 3492-3501. https://doi.org/10.1128/JCM.00957-17
Zhou, Bin ; Deng, Yi Mo ; Barnes, John R. ; Sessions, October M. ; Chou, Tsui Wen ; Wilson, Malania ; Stark, Thomas J. ; Volk, Michelle ; Spirason, Natalie ; Halpin, Rebecca A. ; Kamaraj, Uma Sangumathi ; Ding, Tao ; Stockwell, Timothy B. ; Salvatore, Mirella ; Ghedin, Elodie ; Barr, Ian G. ; Wentworth, David E. / Multiplex reverse transcription-PCR for simultaneous surveillance of influenza A and B viruses. In: Journal of Clinical Microbiology. 2017 ; Vol. 55, No. 12. pp. 3492-3501.
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