Establishment of Detection Method for R292K and N294S Mutation Sites in H3N2 Influenza A Virus with TaqMan-MGB Probes

［Abstract］Objective　To establish a detection method to R292K and N294S mutation sites in H3N2 influenza A virus with TaqMan-MGB probes. Method　NA gene sequences of 23 A（H3N2） influenza strains were acquired from NCBI database and lab isolations. According to sequence alignment results， specific TaqMan-MGB probes and primers were designed for R292K and N294S mutation sites in multiple realtime PCR method. The sensitivity was detected using the ten times dilution method. Clinical specimens， other subtypes of influenza and other common respiratory viruses were used to verify the specificity of the method．Results　The designed probes could detect 107 （1000 copies） dilution of the corresponding template. Other subtypes of influenza virus and common respiratory viruses showed no cross reaction. Through the detection of 57 strains isolated from annual survey， no drug resistance mutations were found. Conclusions　In this study，the A（H3N2） subtype influenza virus R292K and N294S mutation site TaqMan-MGB probe detection method bas been established. It is proved that the primer probe has high sensitivity and specificity with particular application potential.