Development of a real-time fluorescence quantitative RT-PCR assay to detect respiratory syncytial virus B

  Objective   To develop a rapid real-time RT-PCR assay for early detection of human respiratory syncytial virus(RSV)B and quantification of the virus.

  Methods   One pair of primer and one TaqMan-MGB(minor groove binder) fluorogenic probe were designed from the conservative region of N gene of RSVB.Optimized reactive system and thermal cycle conditions were set up.Specificity, sensitivity and repeatability of the method were evaluated.The standard curve and virus quantitative andlysis model were constructed using the standard plasmids in series dilution.One hundred clinical respiratory specimens were detected by this method.

  Results   This method had no cross reaction with other common respiratory virus.The sensitivity was 1 Tissue Culture Infectious Dose₅₀/ml(TCID₅₀/ml).18% of the clinical specimens were positive of RSV B.

  Conclusion   A rapid, sensitive an specific real-time R T-PCR assay to detect RSVB type is established which is suitable for early diagnosis and virus quantification.