Key result
Single-reaction multiplex RT-PCR successfully amplifies critical influenza A and B segments for next-generation sequencing.
Why the study?
Rapid and high-throughput sequencing of influenza A and B viruses remains challenging due to sequence diversity and viral evolution dynamics.
The developed multiplex RT-PCR method enables rapid, high-throughput sequencing of critical genomic segments of seasonal influenza A and B viruses regardless of viral type, subtype, or lineage.
Single-reaction influenza sequencing may accelerate genomic surveillance; leaves open clinical translation and outcome impact.
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.
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Zhou et al. (2017) studied Influenza A and B. Single-reaction influenza A/B virus multiplex reverse transcription-PCR (RT-PCR) was evaluated on Amplification of critical genomic segments (HA, NA, and M) for next-generation sequencing. A single-reaction multiplex RT-PCR method successfully amplified critical genomic segments of seasonal influenza A and B viruses for next-generation sequencing across thousands of specimens.
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