title: [Establishment of a One-Step Real-Time RT-PCR Method for the Detection of Venezuelan Equine Encephalitis Virus].
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.
authors.forename: Shasha
authors.initials: S
authors.lastname: Qian
authors.forename: Biao
authors.initials: B
authors.lastname: He
authors.forename: Zhongzhong
authors.initials: Z
authors.lastname: Tu
authors.forename: Huancheng
authors.initials: H
authors.lastname: Guo
authors.forename: Changchun
authors.initials: C
authors.lastname: Tu
mesh_terms: D002681:China; D017931:DNA Primers / Q000235:genetics; D004666:Encephalitis Virus, Venezuelan Equine / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D004685:Encephalomyelitis, Venezuelan Equine / Q000821:virology*; D006801:Humans; D012367:RNA, Viral / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
chemical_list: D017931:DNA Primers; D012367:RNA, Viral