兔出血症病毒TaqMan荧光定量PCR检测方法的建立  

Establishment of TaqMan real-time PCR for the detection of rabbit hemorrhagic disease virus

作　　者：蒙正群 李桂黎 周丽军[1] 冷依伊 罗怡琳[1] 刘亚东[1] 王印[1,3] 姚学萍[1] 杨泽晓[1] 

机构地区：[1]四川农业大学动物医学院,四川成都611130 [2]四川省动物疫病预防控制中心,四川成都610041 [3]动物疫病与人类健康四川省重点实验室,四川成都611130

出　　处：《中国兽医科学》2017年第6期694-700,共7页Chinese Veterinary Science

基　　金：国家自然科学基金项目(31402222);"十二五"国家科技支撑计划项目(2013BAD12B04);四川省科技支撑计划项目(2016N20002)

摘　　要：为建立快速检测兔出血症病毒(RHDV)的TaqMan荧光定量PCR方法,根据RHDV VP60基因保守序列设计1对特异性引物和1条探针,进行条件优化,检测重复性、敏感性和特异性,建立了RHDV TaqMan荧光定量PCR方法。结果显示,标准品浓度在2.8×10~6~2.8×10~2 copies/L范围内具有良好的线性关系,最低可检测到2.8×10~1copies/L的标准品阳性质粒,变异系数小于2%。结果表明,建立的TaqMan荧光定量PCR方法的特异性、敏感性、重复性均达到试验设计要求,能快速检测临床样品中的RHDV,为我国RHDV的检测及其定量检测的相关研究提供了一种可靠的技术工具。To establish TaqMan real-time PCR for detecting rabbit hemorrhagic disease virus（RHDV） rapidly,a pair of specific primers and a probe were designed based on the conservative gene VP60 of RHDV. The RHDV TaqMan real-time PCR assay was established by the conditions optimization,repeatabili- ty,sensitivity and specificity test. ln results,standards had good liner relations in the concentra-tion of 2.8×10^6~2.8×10^2copies/μL,the limited detection content was 2.8×10^1copies/μL and the va- riation coefficient was less than 2%.In conclusion,the specificity,sensitivity and repeatability of the method satisfy the experimental design requests,and the method can rapidly detect the clinical RHDV samples,providing a reliable technical tool for detection and the quantitative of RHDV in our country.

关 键 词：兔出血症病毒 TAQMAN 荧光定量PCR 

分 类 号：S852.659.6[农业科学—基础兽医学]