Development and evaluation of a RT-qPCR assay for fast and sensitive rabies diagnosis

Rabies virus is endemic to Russia, among other countries. It is therefore critical to develop a high-quality and high-precision diagnostic procedure for the control and prevention of infection. The main objective of the research presented here was to develop a reliable RT-qPCR assay for rabies diagn...

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Bibliographic Details
Published in:Diagnostic microbiology and infectious disease 2018-01, Vol.90 (1), p.18-25
Main Authors: Dedkov, V.G., Deviatkin, A.A., Poleshchuk, Е.М., Safonova, M.V., Blinova, E.A., Shchelkanov, M. Yu, Sidorov, G.N., Simonova, E.G., Shipulin, G.A.
Format: Article
Language:English
Subjects:
Diagnostics
Rabies virus
RT-qPCR
Russia
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Summary:Rabies virus is endemic to Russia, among other countries. It is therefore critical to develop a high-quality and high-precision diagnostic procedure for the control and prevention of infection. The main objective of the research presented here was to develop a reliable RT-qPCR assay for rabies diagnostics. For this purpose, a RABV strains from various biological and geographical origins were used. In addition, rabies-positive and rabies-negative samples, as well as nucleic acids from other viruses and DNA extracted from the brain tissues of mice, dogs, cats, bats and humans, were studied using the developed assay. The analytical sensitivity of the assay, as assessed using armored recombinant positive control dilutions, was 103 copies/ml, and the sensitivity measured using characterized strains was between 0.1 LD50/ml and 1.0 LD50/ml. A broad range of RNA from RABV strains circulating in different regions of Russia, as well as RNA from RABV-positive primary brain samples from 81 animals and two humans, was detected using the developed assay. No false-positive or false-negative results were obtained. Given that high analytical and diagnostic sensitivities and a high specificity were verified for this assay, it has high potential as a screening test that may be suitable for the epizootiological monitoring of animals and for the fast postmortem diagnosis of rabies. •Reliable qRT-PCR assay for RABV detection was developed and evaluated•Analytical sensitivity of the assay was 103 copies/ml, and the sensitivity measured using characterized strains was between 0.1 LD50/1ml and 1.0 LD50/1ml.•Diagnostic sensitivity and specificity were both 100%
ISSN:0732-8893
1879-0070
DOI:10.1016/j.diagmicrobio.2017.09.009