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.
出处
《中国兽医科学》
CAS
CSCD
北大核心
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)
