Establishment of a One-Step Real-Time RT-PCR Method for the Detection of Venezuelan Equine Encephalitis Virus.
- Author:
Shasha QIAN
;
Biao HE
;
Zhongzhong TU
;
Huancheng GUO
;
Changchun TU
- Publication Type:Journal Article
- MeSH:
China;
DNA Primers;
genetics;
Encephalitis Virus, Venezuelan Equine;
classification;
genetics;
isolation & purification;
Encephalomyelitis, Venezuelan Equine;
virology;
Humans;
RNA, Viral;
genetics;
Reverse Transcriptase Polymerase Chain Reaction;
methods
- From:
Chinese Journal of Virology
2015;31(2):107-113
- CountryChina
- Language:Chinese
-
Abstract:
Venezuelan equine encephalitis (VEE) is a zoonotic disease caused by the Venezuelan equine encephalitis virus (VEEV) complex. This disease has not yet been reported in China, and it is therefore essential to establish a rapid and accurate method for detection of the virus in order to prevent and control this disease. In this study, a one-step real-time quantitative RT-PCR method was developed for the detection of the VEEV complex. A pair of specific primers and a Taqman probe were designed corresponding to a conserved region of the VEEV gene nspl, allowing the detection of all known strains of different sub- types of the virus. Using RNA synthesized by in vitro transcription as template, the sensitivity of this method was measured at 3.27 x 10(2) copies/microL. No signal was generated in response to RNA from Chikungunya virus (CHIKV), nor to RNA encoding the nsp1 fragment of Eastern equine encephalitis virus (EE-EV) or Western equine encephalitis virus (WEEV), all of which belong to the same genus as VEEV. This indicates that the method has excellent specificity. These results show that this one-step real-time quantitative RT-PCR method may provide an effective tool for the detection of VEEV in China.
