Establishment of a fluorescent quantitative PCR detection method for rabies virus and preparation of RNA positive controls

VernacularTitle:狂犬病病毒实时荧光定量PCR检测方法的建立和阳性对照的制备
Author: Yun-Long WANG ¹ ; Rui-Hong ZHAO ; Zhi-Tao LI ; Yu-Lin LI ; Guo-Qiang WANG ; Fei SHEN
Author Information

1. 郑州职业技术学院
Keywords: Polymerase chain reaction; Rabies Virus; Virus-Like Particles
From: Chinese Journal of Experimental and Clinical Virology 2008;22(6):504-506
CountryChina
Language:Chinese
Abstract: 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-reslstant virus-like particles for RV positive controls; Results Rnase-resistant vires-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/μl. Conclusion Estabhshed 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.