Objective: To establish a molecular diagnostic method for rabies virus(RV) based on TaqMan PCR.
Methods: BaseD on the rabies virus nucleoprotein gene sequences published in GenBank, RV specific primers and probe were designed by Primer Premier 5.0. The primers and probe were optimized and the sensitivity, specificity,and reproducibility of the system were tested. Quantitative standard curve of RV TaqMan PCR was established. Some RV samples were detected using this system.
Results: The optimized primers and probe were 0.6 micromol/L and 0.2 micromol/L. Reproducibility test showed that coefficient variables were all less than 5% in 4 different system. Quantification standard curve based on the genomic copy was drawn. RV detection using the established method proved that TaqMan PCR was more sensitive and easier performed than traditional RT-PCR.
Conclusion: TaqMan PCR for RV detection had been established, which was more sensitive and specific than the general RT-PCR.