Establishment and application of a TaqMan fluorescence quantitative PCR method for typing influenza B virus
In order to establish a rapid and accurate method for genotyping influenza B virus,the HA gene sequences of Victoria lineage and Yamagata lineage of influenza B virus were downloaded from GISAID and used select the conserved regions for synthesis of a pair of primers and two fluo-rescent probes.Fragments of HA gene of Victoria lineage and Yamagata lineage were amplified u-sing the RNA of Influenza B virus strain preserved in the laboratory and used as a template.The recombinant plasmid standards pUC57-Vic and pUC57-Yama were constructed and identified by PCR and sequencing as plasmid standards.A real-time fluorescence quantitative PCR genotyping method for influenza B virus was preliminarily established.The specificity results showed that the method specifically identified and detected the RNA of Influenza B/Victoria and B/Yamagata Line-ages with no cross-reaction with the RNA/DNA of influenza A virus,EBV,ADV and others.The sensitivity of the detection method was 333 copies/μL and 2 940 copies/μL.The intra-lot CV and inter-lot CV repeatability tests was less than 2％.These data suggested that the established detec-tion method had good specificity,high sensitivity,and good repeatability.The method was used to detect 10 throat swabs of influenza-like cases with conventional reverse transcription PCR(RT-PCR),and the results were consistent(3/10).The fluorescence quantitative PCR method estab-lished in this study can detect and identify influenza B virus infection and provide a more rapid and accurate quantitative detection method for the identification of influenza B virus Victoria lineage and Yamagata lineage.
influenza B virusTaqMan fluorescent quantitative PCRhemagglutinin(HA)
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