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Updated: Jun 22, 2026
Evaluation of a Universal Nested Reverse Transcription Polymerase Chain Reaction for the Detection of Lyssaviruses
Published on: May 2, 2019
[Establishment of a fluorescent quantitative PCR detection method for rabies virus and preparation of RNA positive
Yun-long Wang1, Rui-hong Zhao, Zhi-tao Li
1College of Life Sciences, Henan Normal University, Xinxiang 453007, China. biowyl@126.com
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.
Conclusion:
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.
