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
Central Research Institute for Epidemiology, Moscow, Russia; RAMS Institute of Occupational Health, Moscow, Russia.
Chumakov Institute of Poliomyelitis and Viral Encephalitides, Moscow, Russia; RAMS Institute of Occupational Health, Moscow, Russia.
Diagn Microbiol Infect Dis. 2018 Jan;90(1):18-25. doi: 10.1016/j.diagmicrobio.2017.09.009. Epub 2017 Sep 20.
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 10 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.
狂犬病病毒在俄罗斯等国家呈地方性流行。因此，开发一种高质量、高精度的诊断方法对于控制和预防感染至关重要。本文所开展研究的主要目的是开发一种用于狂犬病诊断的可靠逆转录定量聚合酶链反应（RT-qPCR）检测方法。为此，使用了来自不同生物学和地理来源的狂犬病病毒（RABV）毒株。此外，还使用所开发的检测方法对狂犬病阳性和阴性样本，以及来自其他病毒的核酸和从小鼠、狗、猫、蝙蝠及人类脑组织中提取的DNA进行了研究。使用经修饰的重组阳性对照稀释液评估，该检测方法的分析灵敏度为10拷贝/毫升，使用特征明确的毒株测得的灵敏度在0.1半数致死量（LD50）/毫升至1.0 LD50/毫升之间。使用所开发的检测方法检测到了俄罗斯不同地区流行的多种狂犬病病毒毒株的RNA，以及来自81只动物和两名人类的狂犬病病毒阳性原发性脑样本的RNA。未获得假阳性或假阴性结果。鉴于该检测方法已验证具有高分析灵敏度和诊断灵敏度以及高特异性，它作为一种筛查试验具有很高的潜力，可能适用于动物的动物流行病学监测和狂犬病的快速死后诊断。
