牛病毒性腹泻病毒抗体液相芯片高效检测方法的建立  

Establishment of bovine virtal diarrhea virus antibody detection method by xMAP technology

作　　者：王群 郑小龙 姜帆 马萍萍 于业锋 辛学谦 张丽丽 梁成珠 岳志芹 WANG Qun;ZHENG Xiao-long;JIANG Fan;MA Ping-ping;YU Ye-feng;XIN Xue-qian;ZHANG Li-li;LIANG Cheng-zhu;YUE Zhi-qin(Technology Center of Qingdao Customs,Qingdao 266002,China)

机构地区：[1]青岛海关技术中心,山东青岛266002

出　　处：《中国兽医科学》2022年第10期1268-1272,共5页Chinese Veterinary Science

基　　金：“十三五”国家重点研发计划项目(2016YFD050110401)

摘　　要：为建立一种牛病毒性腹泻病毒(BVDV)血清抗体快速检测的方法,本研究选取BVDV-E2蛋白与荧光微球偶联,建立液相芯片检测技术。首先将BVDV-E2蛋白与已经羧基化处理的荧光微球偶联后与无特定病原血清样本反应,添加检测抗体并利用分析系统检测微球荧光信号建立方法,通过测定无特定病原血清样本确定检测阈值。同时,本试验对液相芯片的最佳蛋白浓度、重复性、一致性等进行了检测。结果显示,本试验建立的液相芯片技术中值荧光强度(MFI)阈值为9347.5,最佳蛋白结合浓度为11μg/1.25×10^(6)个微球,批间和批内变异系数均小于10%,表明该检测方法重复性较好。经过比对发现,本试验建立的方法灵敏度要高于ELISA方法。采用这两种方法对不同血清进行检测,发现两方法的检测一致性为91.6%,且重复性良好。上述研究结果表明,该液相芯片检测技术的建立为BVDV的口岸快速检测探索出一种新的方法。To develop a new rapid detection method for Bovine Viral Diarrhea virus(BVDV),BVDV antibody detection was developed by multi analyze profiling solve for unknow x technology(x MAP).In this paper,BVDV E2 protein was coupled with carboxylated fluorescent microspheres to establish the x MAP technology.Firstly,BVDV-E2 protein was coupled with fluorescent microspheres and reacted with serum samples.After that,the fluorescent signal of microspheres was detected after combined with biotinylated detection antibody.Then the threshold was measured based on median fluorescence intensity(MFI).Furthermore,the optimal concentration of binding protein,the repeatability and consistency of the method were verified.The results showed that the MFI detection threshold was 9347.5.The optimal protein concentration was 11μg/1.25×10^(6)microspheres.The coefficients of variation within and between batches were 9.62%and represent a good repeatability.Meanwhile the serum samples were detected by x MAP and ELISA to detect the consistency,the results showed that the consistency between the two methods were 91.6%.This paper established a high-throughput detection method of BVDV using liquid chip detection technology,and provides a new method for the serological detection in import and export port.

关 键 词：牛病毒性腹泻病毒 抗体检测 液相芯片 酶联免疫吸附试验 荧光微球 

分 类 号：S852.653[农业科学—基础兽医学]