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Multiplex Reverse Transcription-PCR for Simultaneous Surveillance of Influenza A and B Viruses

2017; American Society for Microbiology; Volume: 55; Issue: 12 Linguagem: Inglês

10.1128/jcm.00957-17

1098-660X

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

Viral Infections and Immunology Research

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 (RT-PCR) 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.