蜜蜂慢性麻痹病毒荧光定量RT-PCR检测方法的建立  

Development of a Real-Time RT-PCR Method for Detection of Chronic Bee Paralysis Virus

作　　者：张体银 王武军 林素洁 张志灯 李宋钰 于师宇 ZHANG Tiyin;WANG Wujun;LIN Sujie;ZHANG Zhideng;LI Songyu;YU Shiyu(Fujian Provincial Key Laboratory of Inspection and Quarantine Technology Research,Technology Center of Fuzhou Customs,Fuzhou 350003,China)

机构地区：[1]福州海关技术中心福建省检验检疫技术研究重点实验室,福州350003

出　　处：《中国动物传染病学报》2024年第3期72-78,共7页Chinese Journal of Animal Infectious Diseases

基　　金：海关总署科研项目(2021HK161);福建省对外合作重点项目(2021I0029);福州海关科研项目(FK2020-21)。

摘　　要：为建立慢性蜜蜂麻痹病毒(CBPV)快速诊断方法,本研究根据CBPV RNA依赖RNA聚合酶(RdRp)基因保守区设计特异性引物和TaqMan探针,建立了荧光定量RT-PCR检测方法。结果显示,以构建的重组质粒为标准品建立的TaqMan荧光定量PCR方法,标准曲线具有良好的线性关系,线性相关系数达0.998;该方法最低检出限为10拷贝/μL,与蜜蜂急性麻痹病毒等常见蜜蜂病毒无交叉反应,具有良好的灵敏性和特异性;组内和组间变异系数分别低于0.5%和2%,具有较好的稳定性。本研究建立的CBPV荧光定量RT-PCR检测方法,可用于实验室检测、流行病学调查和疫情监测。To establish a rapid diagnostic method for chronic bee paralysis virus(CBPV),a fluorogenic TaqMan real-time PCR assay was developed by designing specific primers and TaqMan probes according to the RNA dependent RNA polymerase(RdRp)gene sequence of CBPV in this study.The results showed that the real-time RT-PCR method had a good linear relation,and linear correlation coefficient R^(2) was 0.998.The sensitivity limit was 10 copies/μL and there was no cross-reaction with other Honey bee virus,which indicated that the method had good sensitivity and specificity.The coefficients of variation(CV)for intra-assay and inter-assay repeatability were less than 0.5%and 2%,respectively,showing that the method had good stability.The real-time RT-PCR method for CBPV developed in this study might be used for laboratory testing,epidemiological investigation and epidemic monitoring.

关 键 词：蜜蜂慢性麻痹病毒 荧光定量RT-PCR RNA依赖RNA聚合酶 

分 类 号：S895.234[农业科学—特种经济动物饲养]