[Establishment of TaqMan PCR detection method for rabies virus].
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.


CONCLUSIONS
TaqMan PCR for RV detection had been established, which was more sensitive and specific than the general RT-PCR.
Zhang
Qiang
Q
Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
Tang
Qing
Q
Liu
Wei-bin
WB
Li
Hao
H
Liang
Guo-dong
GD
D016133:Polymerase Chain Reaction / Q000379:methods*; D011818:Rabies / Q000175:diagnosis*; D011820:Rabies virus / Q000235:genetics*; D015203:Reproducibility of Results; D019914:Taq Polymerase
D019914:Taq Polymerase