Establishment of a fluorescent quantitative PCR detection method for rabies virus and preparation of RNA positive controls / 中华实验和临床病毒学杂志

ABSTRACT

<p><b>OBJECTIVE</b>Establish the fluorescent quantitative RT-PCR detection method for rabies virus (RV) and construct RNase-resistant virus-like particles as positive controls.</p><p><b>METHODS</b>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;</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>