羊痘病毒属病毒通用Cycleave PCR检测方法的建立  

Development of a Cycleave PCR for Detecting Capripoxvirus

作　　者：南文龙[1] 巩明霞[1] 陆游 李金明[1] 王永杰 哈达 吴发兴[1] 李林[1] 王英丽[1] 陈义平[1] 吴晓东[1] Nan Wenlong;Gong Mingxia;Lu You;Li Jinming;Wang Yongjie;Ha Da;Wu Faxing;Li Lin;Wang Yingli;Chen Yiping;Wu Xiaodong(China Animal Health and Epidemiology Center,Qingdao,Shandong 266032,China;Mongolia Autonomous Region Center for Animal Disease Prevention and Control,Hohhot,Inner Mongolia 010020,China;Xilinhot League Center for Animal Disease Prevention and Control,Xilinhot,Inner Mongolia 026099,China)

机构地区：[1]中国动物卫生与流行病学中心,山东青岛266032 [2]内蒙古自治区动物疫病预防控制中心,内蒙古呼和浩特010020 [3]锡林郭勒盟动物疫病预防控制中心,内蒙古锡林浩特026099

出　　处：《中国动物检疫》2022年第8期107-111,共5页China Animal Health Inspection

摘　　要：为快速特异检测羊痘病毒属病毒(CaPVs),比较了牛结节性皮肤病病毒(LSDV)、山羊痘病毒(GTPV)和绵羊痘病毒(SSPV)的全基因组序列,选择保守区域设计特异性引物和探针,建立了一种CaPVs通用的Cycleave PCR检测方法。特异性结果显示,该方法在40 min内即可检出CaPVs,与口蹄疫病毒、羊口疮病毒、牛病毒性腹泻病毒、牛传染性鼻气管炎病毒、蓝舌病病毒均无交叉反应;灵敏性结果显示,最低检出限为4.98 copies/μL;重复性结果显示,批内和批间重复变异系数均小于2%。对131份临床样本进行检测发现,该方法与世界动物卫生组织(WOAH)推荐的常规PCR方法相比,敏感性为100%,特异性为91.55%,总符合率为95.42%。以上结果表明,本研究建立的Cycleave PCR检测方法特异、敏感、重复性好,且操作简便,适用于CaPVs的高通量、快速检测。In order to detect Capripoxvirus(CaPVs)rapidly and specifically,the whole genomes of lumpy skin disease virus(LSDV),goatpox virus(GTPV)and sheeppox virus(SSPV)were compared,then specific primers and probes were designed based on the conserved region to develop a Cycleave PCR assay for CaPVs.The results showed that,for the specificity,CaPVs could be detected by the assay within 40 min,and no cross reaction was observed for foot-and-mouth disease virus(FMDV),orf virus(ORFV),bovine viral diarrhea virus(BVDV),infectious bovine rhinotracheitis virus(IBRV)and bluetongue virus(BTV);for the sensitivity,the minimum detection limit was 4.98 copies/μL;and for the repeatability,the coefficient of repeated variation of intra batch and inter batch was<2%.It was found that,based on the test results of 131 clinical samples,compared with the routine PCR assays recommended by World Organization for Animal Health(WOAH),the sensitivity,specificity and total coincidence rate of the assay were 100%,91.55%and 95.42%,respectively.In conclusion,the established Cycleave PCR assay,with the help of its high specificity,sensitivity,repeatability and simple operation,was appropriate for high-throughput and rapid detection of CaPVs.

关 键 词：羊痘病毒属病毒 Cycleave PCR 检测 

分 类 号：S855.3[农业科学—临床兽医学]