Real-time fluorescent quantitative RT-PCR methods for detection of Avalon virus and Hughes virus
10.3760/cma.j.cn112866-20200826-00231
- VernacularTitle:阿瓦朗病毒和休斯病毒实时荧光RT-PCR检测方法研究
- Author:
Shanshan DU
1
;
Aqian LI
;
Xiaoxia HUANG
;
Yang LIU
;
Qin WANG
;
Chuan LI
;
Mifang LIANG
;
Dexin LI
;
Shiwen WANG
;
Jiandong LI
Author Information
1. 中国疾病预防控制中心病毒病预防控制所 国家卫生健康委员会生物安全重点实验室，北京 102206
- Keywords:
Nairoviruses;
Quantitative real-time PCR;
Avalon virus;
Hughes virus
- From:
Chinese Journal of Experimental and Clinical Virology
2021;35(1):111-115
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish real-time fluorescent quantitative RT-PCR method for detection of Avalon virus (AVAV) and Hughes virus (HUGV), two Nairoviruses in the family of Nairoviridae. Methods:The genomic sequences of the two viruses published in the international public database were collected, collated, compared and analyzed to define the detection targets, and the viral specific primers and probes were designed accordingly. Real-time fluorescent quantitative RT-PCR detection method were established, and the operating detection procedures were optimized using simulated samples prepared using in vitro transcription assay, other virus infected samples, virus strains and normal human blood samples. The detection limit, specificity and repeatability of the method were evaluated.Results:The real-time fluorescent quantitative RT-PCR method could effectively amplify and detect AVAV and HUGV viral target RNA with detection limits of about 20 copies/μl and 70 copies/μl, respectively. No nonspecific amplification was found in the samples of Kyasanur forest disease virus, influenza B virus BV and BY, influenza A virus H3N2, yellow fever virus, Japanese encephalitis virus, Crimean-Congo hemorrhagic fever virus, SFTS virus, nairobi sheep disease virus and Tahyna virus. There was no cross reaction between the two nairoviruses. The coefficient of variation was within 2% by repeated comparative analysis.Conclusions:The real-time fluorescent quantitative RT-PCR method for detection of AVAV and HUGV might be used for screening of humans, vectors and host animal samples for rapid detection of related pathogens.
