山羊痘病毒PCR检测方法的建立及应用  

Development and Application of PCR for Detection of Goat Pox Virus

作　　者：白艳艳[1] 张斌 郝玉青 高艳[1] 刘万华[1] 白崇生 刘健鹏[1] 韩斌[1] 杨德全[3] 

机构地区：[1]榆林市动物疫病预防控制中心,陕西榆林719000 [2]榆林市动物卫生监督所,陕西榆林719000 [3]上海市动物疫病预防控制中心,上海201103

出　　处：《动物医学进展》2017年第9期10-13,共4页Progress In Veterinary Medicine

基　　金：陕西省农业科技创新转化资金项目(NYKJ-2015-023);陕西省科技统筹项目(2015KTTSNY04-04)

摘　　要：为了更快、更准确的诊断山羊痘(Goat pox),根据GenBank上已公布的山羊痘病毒(GTPV)的基因序列,针对p32基因的保守序列设计并合成一对能特异性扩增山羊痘病毒的引物,扩增产物大小约为983bp。经过反应条件的优化,建立了山羊痘病毒PCR检测方法,对所建立的PCR反应体系的特异性和灵敏性进行了评价,并用此方法对9份临床样品进行了检测。结果显示,该诊断方法与3种非羊痘病毒不发生交叉反应。该方法最低浓度检测限为0.4pg。在检测的临床样品中,GTPV阳性有3份。结果表明,所建立的方法具有良好的特异性和敏感性,适合临床诊断应用。The purpose of this study was to develop a faster and more accurate diagnostic method. Based on the published reference strains of goat pox virus（GTPV） in GenBank,a pair of primers for p32 gene se-quence of GPTV specific amplification were designed and synthesized. The amplified product was approxi-mately 983 bp. After the optimization of reaction condition, a PCR method for detection of GTPV was es-tablished and an evaluation on the specificity and sensitivity of reaction system for this method was conduc-ted with detecting 9 clinical samples by this method. The result indicated that this diagnostic method did not generate cross reaction with three kinds of non-capripoxviruses. The minimum concentration of PCR is 0. 4 pg. In the clinical samples detected, the number of positive GTPV is 3. The result showred that the method has good specificity and sensitivity, which can be suitable for clinical diagnosis.

关 键 词：山羊痘 聚合酶链反应 敏感性 特异性 

分 类 号：S852.659.1[农业科学—基础兽医学] S854.43[农业科学—兽医学]