中国兽医科学2024，Vol.54Issue(8)：1002-1009,8.DOI:10.16656/j.issn.1673-4696.2024.0138
基于CA-TM融合蛋白的梅迪-维斯纳病毒胶体金免疫层析检测方法的建立与应用
Establishment and application of colloidal gold immunochromatographic assay for detecting Maedi-Visna virus based on CA-TM fusion protein
摘要
Abstract
The purpose of this study was to establish a rapid and accurate colloidal gold im-munochromatographic method for the detection of Maedi-Visna virus(MVV)antibodies.The recombinant CA-TM fusion protein(rCA-TM)labeled detection line which has been authorized by our laboratory.The quality con-trol line was labeled with chicken IgY antibody,and r-SPG and chicken IgY were used for colloidal gold labeling.The colloidal gold immunochromatography method for rapid detection of MVV was established after optimization of various conditions,and the detection effect was evaluated by specificity,sensi-tivity,stability and clinical application.In result,we successfully expressed,purified and obtained the colloidal gold immunochromatographic assay established by rCA-TM;the specificity test results of colloidal gold immunochromatography showed that it could not detect other common sheep pathogens such as foot-and-mouth disease virus,Peste des petits ruminant virus,Mycoplasma ovis and Brucella except MVV,and results of sensitivity test showed that MVV-positive serum with a minimum dilution of 2 400 times could be detected.The stability test results showed that the test strip still had good detection performance after 6 months of sealed storage at room temperature,and the MVV colloidal gold detection method and the imported commercial MVV antibody ELISA detection kit widely used in MVV detection were used to detect 40 serum samples of suspected cases.The results showed that 17 MVV antibody positive sera and 23 negative sera were detected by colloidal gold detection.The imported commercial MVV anti-body ELISA detection kit detected 16 positive sera and 24 negative sera,with an overall coincidence rate of 97.5％.In summary,this experiment successfully established a rapid detection method of MVV colloidal gold immunochromatography.The test strip developed by this method has good performance,is easy to store,and is suitable for on-site rapid detection and batch detection.It is of great signifi-cance for the monitoring,prevention and control of MVV,and provides a new detection scheme for the pu-rification of MVV in Inner Mongolia.关键词
梅迪-维斯纳病毒/原核表达/融合蛋白/胶体金/免疫层析Key words
Maedi-Visna virus/prokaryotic expression/fusion protein/colloidal gold/immunochro-matography分类
农业科技引用本文复制引用
陈思旭,包磊,李慧萍,张良,刘淑英..基于CA-TM融合蛋白的梅迪-维斯纳病毒胶体金免疫层析检测方法的建立与应用[J].中国兽医科学,2024,54(8):1002-1009,8.基金项目
国家自然科学基金项目(32072819,32360863) （32072819,32360863）
内蒙古科技重大专项计划项目(2021ZD0010) （2021ZD0010）
内蒙古草原英才创新团队项目(20151031) （20151031）
内蒙古教育厅兽医基础与牛羊疫病防控技术研发创新团队项目(NMGIRT2412) （NMGIRT2412）
内蒙古自治区研究生科研创新项目(B20210174Z) （B20210174Z）
