基于SYBR GreenⅡ的BVDV NS3基因实时荧光定量PCR检测方法的建立与应用  

Establishment and Application of qRT-PCR Assay for BVDV NS3 GeneBased on SYBR Green Ⅱ

作　　者：李阳[1,2] 张世勋 赫鸣睿[1] 刘珊珊 岳山[1] 刘宇[1,2] 朱战波[1,2] Li Yang;Zhang Shixun;He Mingrui;Liu Shanshan;Yue Shan;Liu Yu;Zhu Zhanbo(College of Animal Science and Veterinary Medicine,Heilongjiang Bayi Agricultural University,Daqing 163319;Engineering Research Center of Prevention and Control of Cattle Diseases)

机构地区：[1]黑龙江八一农垦大学动物科技学院,大庆163319 [2]黑龙江省牛病防控工程技术研究中心

出　　处：《黑龙江八一农垦大学学报》2025年第2期23-31,共9页journal of heilongjiang bayi agricultural university

基　　金：黑龙江省“揭榜挂帅”科技攻关项目,2023ZXJ02B03。

摘　　要：为了建立基于SYBR GreenⅡ的牛病毒性腹泻病毒(BVDV)NS3基因实时荧光定量PCR(qRT-PCR)检测方法,设计并筛选了BVDV NS3基因的qRT-PCR引物,建立了BVDV NS3 qRT-PCR检测方法,并利用CFX96和猪警-2000 qRT-PCR仪检测了临床牛血清样本。结果显示,筛选出1对特异性好的引物,CFX96和猪警-2000 qRT-PCR的标准品最小检出值分别为1×10^(1)copies·μL^(-1)和1×10^(2) copies·μL^(-1),特异性和重复性均良好,且临床血清样本的检测结果准确、一致。研究为BVDV检测提供了技术手段。To establish a real-time fluorescent quantitative PCR(qRT-PCR)method for the detection of bovine viral diarrhea virus(BVDV)NS3 gene based on SYBR GreenⅡ,the qRT-PCR primers of BVDV NS3 gene were designed and screened,and the qRT-PCR detection method was established.Clinical bovine serum samples were also detected by CFX96 and pig early warning machine-2000.The results showed that one pair of primers with good specificity was selected.The minimum detectable values of CFX96 and pig early warning machine-2000 qRT-PCR standards were 1×10^(1) copies·μL^(-1) and 1×10^(2) copies·μL^(-1),respectively,with good specificity and repeatability,and the detection results of clinical serum samples were accurate and consistent.It provides technical means for BVDV detection.

关 键 词：牛病毒性腹泻病毒 NS3基因 实时荧光定量PCR SYBR GreenⅡ 

分 类 号：S857.21[农业科学—临床兽医学]