关键词：
帕利亚姆病毒;一步法RT-PCR;血清型鉴别
摘 要：
帕利亚姆血清群病毒(Palyam serogroup virus,PALV)包含多种血清型病毒,我国存在Chuzan virus(CHUV)、Bunyip Creek virus (BCV)和D'Aguilar virus (DAV)三种血清型PALV毒株的流行,目前尚无PALV血清型特异性RT-PCR检测方法的报道。本研究根据获取的中国CHUV、BAV和DAV这三种PALV血清型毒株的Seg-2序列,设计合成了3对特异性引物,建立了PALV血清型特异性一步法RT-PCR检测方法。灵敏度试验结果显示,CHUV、BCV和DAV血清型鉴定引物可分别检测到1.3×10~2 copies、8.0×10~2 copies和5.5×10~2 copies的核酸;特异性试验结果表明,本研究建立的PALV血清型特异性检测方法与蓝舌病病毒(BTV)、流行性出血病病毒(EHDV)、非洲马瘟病毒(ASHV)均无交叉反应。本研究建立的PALV血清型特异性RT-PCR检测方法具有良好的特异性和较高的灵敏度,为开展PALV的诊断与流行病学监测提供了有效的技术手段。
译 名：
Establishment of a serotype-specific RT-PCR for the detection of Palyam serogroup virus
作 者：
LI Zhan-hong;YANG Zhen-xing;ZHANG Jie;XIAO Lei;LIAO De-fang;LI Hua-chun;YANG Heng;Yunnan Tropical and Subtropical Animal Virus Diseases Laboratory,Yunnan Animal Science and Veterinary Institute;College of Veterinary Medicine,Yunnan Agricultural University;
关键词：
Palyam serogroup virus;;one-step RT-PCR;;serotype identification
摘 要：
Palyam serogroup virus(PALV)contains many serotypes,and there are three serotypes of PALV strains,Chuzan virus(CHUV),Bunyip Creek virus(BCV) and D'Aguilar virus(DAV) in China. There was no report about the serotype-specific RT-PCR method for PALV,currently. Three pairs of specific primers were designed based on the seg-2 sequences of Chinese CHUV,BAV and DAV strains. A PALV serotype specific one-step RT-PCR detection assay was established. Sensitivity tests showed that the lowest detection levels of CHUV,BCV and DAV were 1.3 ×10~2 copies,8.0 ×10~2 copies and 5.5 ×10~2 copies by the one-step RT-PCR,respectively. This method had no cross reaction with bluetongue virus(BTV),epidemic hemorrhagic disease virus(EHDV),and african equine plague virus(AHSV). The results indicated that the PALV serotype-specific one step RT-PCR had ideal specificity and high sensitivity and it provided an effective technical means for the diagnosis and epidemiological monitoring of PALV in China.
