蓝舌病病毒8型实时荧光定量RT-PCR检测方法的建立与应用  

Development and Application of a RT-qPCR Assay for Detection Bluetongue Virus Serotype 8

作　　者：林彦星 花群俊 张彩虹 曹琛福 阮周曦 曾少灵 史卫军 黄超华 杨俊兴 花群义 LIN Yan-xing;HUA Qun-jun;ZHANG Cai-hong;CAO Chen-fu;RUAN Zhou-xi;ZENG Shao-ling;SHI Wei-jun;HUANG Chao-hua;YANG Jun-xing;HUA Qun-yi(Inspection and Quarantine Center for Animals and Plants,Shenzhen Customs,Shenzhen,Guangdong,518045,China;Wuding Center for Animal Disease Control and Prevention,Wuding,Yunnan,651601,China)

机构地区：[1]深圳海关动植物检验检疫技术中心,广东深圳518045 [2]云南省武定县动物疫病预防控制中心,云南武定651601

出　　处：《动物医学进展》2020年第8期44-48,共5页Progress In Veterinary Medicine

基　　金：国家重点研发计划项目资助(2017YFD0501805、2017YFD0502304)。

摘　　要：根据蓝舌病病毒8型(BTV-8)VP2基因序列(Seg-2)设计特异性引物和探针,建立了BTV-8实时荧光定量RT-PCR(RT-qPCR)检测方法。结果表明该法特异性强,能准确检测BTV-8核酸,与蓝舌病病毒其他血清型病毒、鹿流行性出血热病毒(EHDV)、牛病毒性腹泻病毒(BVDV)、口蹄疫病毒(FMDV)等病原核酸无交叉反应;最低可检测核酸浓度为2.8×10^3 copies/mL;Ct值组内和组间变异系数(CV)均小于2%。用所建立方法对115份临床样品进行检测,结果均为阴性,与文献报道的方法检测结果一致。建立的RT-qPCR方法可用于BTV-8的临床检测和流行病学调查。In this study,a fluorescent RT-PCR(RT-qPCR)was developed to detect the bluetongue virus serotype 8(BTV-8)using primers and probes specific for the genome segment 2,which encodes the outer shell protein VP2.The results showed that the RT-qPCR assay was specific for BTV-8 and does not cross-react with other serotypes of bluetongue virus,EHDV,BVDV or FMDV.The limit of detection of RT-qPCR for BTV-8 was 2.8×10^3 copies/mL in this test.A total of 115 samples were tested by the RT-qPCR assay and the other assay from reference,and the test results were consistent.The RT-qPCR assay could be used for clinical detection and epidemiological investigation of BTV-8.

关 键 词：蓝舌病病毒8型 实时荧光定量RT-PCR 检测 

分 类 号：S851.33[农业科学—预防兽医学] S852.659.4[农业科学—兽医学]