Format

Send to

Choose Destination
J Clin Microbiol. 2017 Dec;55(12):3492-3501. doi: 10.1128/JCM.00957-17. Epub 2017 Oct 4.

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

Author information

1
Center for Genomics and Systems Biology, Department of Biology, New York University, New York, New York, USA.
2
J. Craig Venter Institute, Rockville, Maryland, USA.
3
World Health Organization Collaborating Centre for Reference and Research on Influenza, The Peter Doherty Institute for Infection and Immunity, Melbourne, Australia yi-mo.deng@influenzacentre.org dwentworth@cdc.gov.
4
Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA.
5
Program in Emerging Infectious Diseases, Duke-NUS Graduate Medical School Singapore, Singapore.
6
World Health Organization Collaborating Centre for Reference and Research on Influenza, The Peter Doherty Institute for Infection and Immunity, Melbourne, Australia.
7
Department of Medicine and Department of Healthcare Policy and Research, Weill Cornell Medical College of Cornell University, New York, New York, USA.
8
College of Global Public Health, New York University, New York, New York, USA.
9
J. Craig Venter Institute, Rockville, Maryland, USA yi-mo.deng@influenzacentre.org dwentworth@cdc.gov.

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 (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.

KEYWORDS:

NGS; RT-PCR; influenza; surveillance

PMID:
28978683
PMCID:
PMC5703814
DOI:
10.1128/JCM.00957-17
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for HighWire Icon for PubMed Central
Loading ...
Support Center