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

tetano

Editor, Senior Moderator
Arch Virol. 2016 Feb 29. [Epub ahead of print]
[h=1]Lineage-specific detection of influenza B virus using real-time polymerase chain reaction with melting curve analysis.[/h] Tewawong N[SUP]1[/SUP], Chansaenroj J[SUP]1[/SUP], Klinfueng S[SUP]1[/SUP], Vichiwattana P[SUP]1[/SUP], Korkong S[SUP]1[/SUP], Thongmee T[SUP]1[/SUP], Theamboonlers A[SUP]1[/SUP], Payungporn S[SUP]2[/SUP], Vongpunsawad S[SUP]1[/SUP], Poovorawan Y[SUP]3[/SUP].
[h=3]Author information[/h]

[h=3]Abstract[/h] 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 10[SUP]2[/SUP] 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.


PMID: 26923928 [PubMed - as supplied by publisher]
 
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