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当前位置: 首页 > 文章 > 牛结节性皮肤病病毒Cycleave PCR检测方法的建立 中国兽医学报 2023 (6) 1145-1148,1174
Position: Home > Articles > 牛结节性皮肤病病毒Cycleave PCR检测方法的建立 Chinese Journal of Veterinary Science 2023 (6) 1145-1148,1174

牛结节性皮肤病病毒Cycleave PCR检测方法的建立

作  者：
南文龙;巩明霞;陆游;李金明;王永杰;哈达;李林;杜秋明;陈义平;吴晓东
单  位：
延边州动物疫病预防控制中心;内蒙古自治区动物疫病预防控制中心;中国动物卫生与流行病学中心;锡林郭勒盟动物疫病预防控制中心
关键词：
牛结节性皮肤病病毒;Cycleave PCR;ORF024基因
摘  要：
将GenBank中牛结节性皮肤病病毒与同属的山羊痘病毒和绵羊痘病毒全基因组进行比较，筛选设计特异性引物和探针，建立Cycleave PCR检测方法。结果显示，建立的方法在40 min内即可特异性地检出牛结节性皮肤病病毒核酸，与山羊痘病毒、绵羊痘病毒、口蹄疫病毒、牛病毒性腹泻/黏膜病病毒、牛传染性鼻气管炎病毒、蓝舌病病毒均无交叉反应，该方法的最低检出限为1.13拷贝/μL。平行检测89份临床样本，与国标荧光定量PCR方法相比，本方法敏感性100.0%,特异性90.0%,总符合率为95.5%。结果表明，本试验建立的Cycleave PCR检测方法敏感、特异且操作简便，适用于牛结节性皮肤病病毒的高通量、快速检测。
作  者：
NAN Wenlong;GONG Mingxia;LU You;LI Jinming;WANG Yongjie;HA Da;LI Lin;DU Qiuming;CHEN Yiping;WU Xiaodong;China Animal Health and Epidemiology Center;Inner Mongolia Autonomous Region Animal Disease Prevention and Control Center;Xilin Gol League Animal Disease Prevention and Control Center;Yanbian Animal Disease Prevention and Control Center;
关键词：
lumpy skin disease virus;;Cycleave PCR;;ORF024 gene
摘  要：
In order to detect lumpy skin disease virus(LSDV) rapidly and specifically, the whole genomes of LSDV,goatpox virus and sheeppox virus in GenBank were compared, and the specific primers and probes were designed, which were then used to establish a Cycleave PCR detection method.The results showed that the Cycleave PCR could specifically detect the nucleic acid of LSDV within 40 min, and had no cross reaction with goatpox virus, sheeppox virus, foot-and-mouth disease virus, bovine viral diarrhea/mucosal disease virus, bovine infectious rhinotracheitis virus and bluetongue virus.The low detection limit was 1.13 copies/μL.To compared with the national standard real-time PCR method, 89 clinical samples were tested in parallel with Cycleave PCR,and the sensitivity, specificity and total coincidence rate were 100.0%,90.0% and 95.5%,respectively.The results indicated that the established Cycleave PCR was sensitive, specific and easy to operate, which was suitable for high-throughput and rapid detection of LSDV.

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