Journal of clinical microbiology · 2017
Multiplex Reverse Transcription-PCR for Simultaneous Surveillance of Influenza A and B Viruses.
СтатьяВалидационное исследованиеНазвание и аннотация: Перевод пока не подготовлен
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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
Аннотация
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.