[Establishment of a fluorescent quantitative PCR detection method for rabies virus and preparation of RNA positive controls].
OBJECTIVE
Establish the fluorescent quantitative RT-PCR detection method for rabies virus (RV) and construct RNase-resistant virus-like particles as positive controls.


METHODS
Analyze the database in GenBank, the probe and the primers were designed in the conservative region of N gene of rabies virus and the method of real-time fluorescent quantitative PCR was obtained; On the basis of MS2 phage, with the technology of gene recombination, prepare the RNase-resistant virus-like particles for RV positive controls;


RESULTS
RNase-resistant virus-like particles were obtained after prokaryotic expression in E. coli. The designed primers and probe were confirmed to be very specific and conservative, and be sensitive to-concentration of 15 copies/microl.


CONCLUSIONS
Established the method of detecting rabies virus by reverse transcription real-time quantitative PCR,obtained the RNase-resistant and no infectivity virus-like particles as positive controls of rabies virus.
Wang
Yun-long
YL
College of Life Sciences, Henan Normal University, Xinxiang 453007, China. biowyl@126.com
Zhao
Rui-hong
RH
Li
Zhi-tao
ZT
Li
Yu-lin
YL
Wang
Guo-qiang
GQ
Shen
Fei
F
D000818:Animals; D015342:DNA Probes; D004279:DNA, Viral; D004285:Dogs; D006801:Humans; D012367:RNA, Viral / Q000032:analysis*; D011818:Rabies / Q000821:virology; D011820:Rabies virus / Q000737:chemistry*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012259:Ribonuclease, Pancreatic / Q000378:metabolism; D019562:Viral Load
D015342:DNA Probes; D004279:DNA, Viral; D012367:RNA, Viral; D012259:Ribonuclease, Pancreatic