title: Applicability of a sensitive duplex real-time PCR assay for identifying B/Yamagata and B/Victoria lineages of influenza virus from clinical specimens.
abstract: Type B influenza virus is one of the major epidemic strains and responsible for considerable mortality and morbidity. Rapidly and accurately identifying different influenza B virus lineages, i.e., B/Yamagata (B/Y) and B/Victoria (B/V), is desirable during the flu season. However, the available rapid techniques lack sensitivity, and the usual methods for identifying influenza viruses require expansion of virus in tissue culture or embryonated hen's eggs. Thus, we developed several sets of primer pairs that were able to detect and distinguish B/Y and B/V in a single real-time PCR assay. Used in conjunction with two sets of specific primers that exhibited purine at 3' end of at least one primer targeting on HA gene of B/Y and B/V lineages allows us to accurately identify approximately 10(2) copies per microliter for B/Y and B/V with intra- and inter-assay coefficient of variation (CV) <4%. When it was used to test 17,765 throat swab specimens obtained in the 2006-2010 influenza surveillance season, this method was comparable to hemagglutination inhibition assay in detection, typing and subtyping of influenza viruses with 100% true-negative (specificity) and 100% true-positive (sensitivity). Taken together, this method provides sensitive and robust tool for routine diagnosis and on-time epidemiological examination for WHO decisions on vaccine composition.
authors: Department of Social Medicine, Public Health School, Tongji Medical College, Huazhong University of Science and Technology, 13 Hangkong Road, Wuhan, 430030, People's Republic of China.
authors: Naixing
authors: N
authors: Zhang
authors: Shisong
authors: S
authors: Fang
authors: Ting
authors: T
authors: Wang
authors: Jianxiong
authors: J
authors: Li
authors: Xiaowen
authors: X
authors: Cheng
authors: Cunyou
authors: C
authors: Zhao
authors: Xin
authors: X
authors: Wang
authors: Xing
authors: X
authors: Lv
authors: Chunli
authors: C
authors: Wu
authors: Renli
authors: R
authors: Zhang
authors: Jinquan
authors: J
authors: Cheng
authors: Hong
authors: H
authors: Xue
authors: Zuxun
authors: Z
authors: Lu
mesh_terms: D017931:DNA Primers / Q000235:genetics; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics; D006801:Humans; D009981:Influenza B virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D007251:Influenza, Human / Q000175:diagnosis* / Q000821:virology*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D010614:Pharynx / Q000821:virology; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods*
chemical_list: D017931:DNA Primers; D019267:Hemagglutinin Glycoproteins, Influenza Virus