title: Lineage-specific detection of influenza B virus using real-time polymerase chain reaction with melting curve analysis.
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 10(2) 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.
authors.affiliation: Center of Excellence in Clinical Virology, Department of Pediatrics, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand.
authors.forename: Nipaporn
authors.initials: N
authors.lastname: Tewawong
authors.affiliation: Center of Excellence in Clinical Virology, Department of Pediatrics, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand.
authors.forename: Jira
authors.initials: J
authors.lastname: Chansaenroj
authors.affiliation: Center of Excellence in Clinical Virology, Department of Pediatrics, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand.
authors.forename: Sirapa
authors.initials: S
authors.lastname: Klinfueng
authors.affiliation: Center of Excellence in Clinical Virology, Department of Pediatrics, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand.
authors.forename: Preeyaporn
authors.initials: P
authors.lastname: Vichiwattana
authors.affiliation: Center of Excellence in Clinical Virology, Department of Pediatrics, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand.
authors.forename: Sumeth
authors.initials: S
authors.lastname: Korkong
authors.affiliation: Center of Excellence in Clinical Virology, Department of Pediatrics, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand.
authors.forename: Thanunrat
authors.initials: T
authors.lastname: Thongmee
authors.affiliation: Center of Excellence in Clinical Virology, Department of Pediatrics, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand.
authors.forename: Apiradee
authors.initials: A
authors.lastname: Theamboonlers
authors.affiliation: Department of Biochemistry, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.
authors.forename: Sunchai
authors.initials: S
authors.lastname: Payungporn
authors.affiliation: Center of Excellence in Clinical Virology, Department of Pediatrics, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand.
authors.forename: Sompong
authors.initials: S
authors.lastname: Vongpunsawad
authors.affiliation: Center of Excellence in Clinical Virology, Department of Pediatrics, Faculty of Medicine, Chulalongkorn University, Bangkok, 10330, Thailand. yong.p@chula.ac.th.
authors.forename: Yong
authors.initials: Y
authors.lastname: Poovorawan
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D002648:Child; D002675:Child, Preschool; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D005814:Genes, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics / Q000276:immunology; D006801:Humans; D007223:Infant; D009981:Influenza B virus / Q000145:classification* / Q000235:genetics* / Q000276:immunology; D007252:Influenza Vaccines / Q000494:pharmacology; D007251:Influenza, Human / Q000175:diagnosis / Q000453:epidemiology / Q000821:virology; D008875:Middle Aged; D017720:Molecular Epidemiology; D009691:Nucleic Acid Denaturation; D015995:Prevalence; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D055815:Young Adult
chemical_list: D004279:DNA, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D007252:Influenza Vaccines