TaqMan-based Real-time RT-PCR Assay for Quick Detection of Influenza A_1 Virus RNA

Objective To establish a TaqMan-based real-time PCR assay for detection of influenza A_1 virus RNA.Methods The specific primers and probes were designed in the conserved region of influenza A_1 virus the HA gene.The primers and probes are selected,the reactive condition was and system optimized to improve the sensitivity,specificity and repetition of the assay.The clinical specimens collected from the patients with acute respiratory tract infection were detected by this assay.Results The specificity of the assay was high and there were no cross reactions with influenza A_3,B,A_5 virus SARS virus,measles virus and the other common respiratory virus.The sensitivity of the assay was 0.1 TCID_(50);Specificity and sensitivity of the rea-time PCR method were compared to the conventional virus isolating using MDCK cells the detection rate of influenza A_1 virus from the throat of the clinical patients,washing specimens by RT-PCR method was much higher than that by conventional method.It took only three hours to extract viral RNA and do the real-time PCR.It had advantages of contamination control,automation and direct virus quantification.Conclusion This TaqMan-based real-time PCR assay was a quick,sensitive and specific tool for molecular diagnosis of influenza A_1 virus.