Establishment of a fluorescent quantitative PCR detection method for rabies virus and preparation of RNA positive controls.
- Author:
Yun-long WANG
1
;
Rui-hong ZHAO
;
Zhi-tao LI
;
Yu-lin LI
;
Guo-qiang WANG
;
Fei SHEN
Author Information
- Publication Type:Journal Article
- MeSH: Animals; DNA Probes; DNA, Viral; Dogs; Humans; RNA, Viral; analysis; Rabies; virology; Rabies virus; chemistry; Reverse Transcriptase Polymerase Chain Reaction; methods; Ribonuclease, Pancreatic; metabolism; Viral Load
- From: Chinese Journal of Experimental and Clinical Virology 2008;22(6):504-506
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVEEstablish the fluorescent quantitative RT-PCR detection method for rabies virus (RV) and construct RNase-resistant virus-like particles as positive controls.
METHODSAnalyze 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;
RESULTSRNase-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.
CONCLUSIONEstablished 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.
