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Lineage-specific detection of influenza B virus using real-time polymerase chain reaction with melting curve analysis

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Abstract

Influenza B viruses comprise two lineages, Victoria (B/Vic) and Yamagata (B/Yam), which co-circulate globally. The surveillance data on influenza B virus lineages in many countries often underestimate the true prevalence due to the lack of a rapid, accurate, and cost-effective method for virus detection. We have developed a real-time PCR with melting curve analysis for lineage-specific differential detection of influenza B virus. By amplifying a region of the hemagglutinin gene using real-time PCR with SYBR Green I dye, B/Vic and B/Yam could be differentiated based on their melting temperature peaks. This method was efficient (B/Vic = 93.2 %; B/Yam 97.7 %), sensitive (B/Vic, 94.6 %; B/Yam, 96.3 %), and specific (B/Vic, 97.7 %; B/Yam, 97.1 %). The lower detection limit was 102 copies per microliter. The assay was evaluated using 756 respiratory specimens that were positive for influenza B virus, obtained between 2010 and 2015. The incidence of influenza B virus was approximately 18.9 % of all influenza cases, and the percentage was highest among children aged 6–17 years (7.57 %). The overall percentage of mismatched influenza B vaccine was 21.1 %. Our findings suggest that real-time PCR with melting curve analysis can provide a rapid, simple, and sensitive lineage-specific influenza B virus screening method to facilitate influenza surveillance.

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Acknowledgments

This work was supported by the National Research University Project, Office of Higher Education Commission (WCU001-HR-57, WCU007-HR-57, and WCU-58-006-HR), the National Research Council of Thailand, the Research Chair Grant from the National Science and Technology Development Agency, Chulalongkorn University Centenary Academic Development Project (CU56-HR01), Ratchadaphiseksomphot Endowment Fund of Chulalongkorn University (RES560530093), the Outstanding Professor of Thailand Research Fund (DPG5480002), Siam Cement Group, and MK Restaurant Company Limited. The 100th Anniversary Chulalongkorn University Fund for doctoral scholarship was awarded to Nipaporn Tewawong.

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Authors and Affiliations

  1. Center of Excellence in Clinical Virology, Department of Pediatrics, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand

    Nipaporn Tewawong, Jira Chansaenroj, Sirapa Klinfueng, Preeyaporn Vichiwattana, Sumeth Korkong, Thanunrat Thongmee, Apiradee Theamboonlers, Sompong Vongpunsawad & Yong Poovorawan

  2. Department of Biochemistry, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand

    Sunchai Payungporn

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  1. Nipaporn Tewawong
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  2. Jira Chansaenroj
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Correspondence to Yong Poovorawan.

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Tewawong, N., Chansaenroj, J., Klinfueng, S. et al. Lineage-specific detection of influenza B virus using real-time polymerase chain reaction with melting curve analysis. Arch Virol 161, 1425–1435 (2016). https://doi.org/10.1007/s00705-016-2802-2

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