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

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国家科技期刊平台
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中文摘要：旨在建立一种快速、准确的梅迪-维斯纳病毒(MVV)抗体的胶体金免疫层析检测方法.利用本实验室的重组CA-TM融合蛋白(rCA-TM)标记检测线,以鸡IgY抗体标记质控线,以重组链球菌蛋白(r-SPG)与鸡IgY用于胶体金标记.经各条件优化建立MVV胶体金免疫层析快速检测方法,并通过特异性、敏感性、稳定性以及临床应用对其检测效果进行评价.结果显示,成功表达、纯化并获得了rCA-TM;成功建立的胶体金免疫层析法特异性试验结果显示不能检测除MVV 以外的如口蹄疫病毒、小反刍兽疫病毒、绵羊肺炎支原体、布鲁菌等绵羊常见病原;敏感性试验结果显示最低可检出2 400倍稀释的MVV阳性血清;稳定性试验结果显示在模拟6个月避光常温密封保存后的试纸条仍具有良好的检测性能.利用建立的MVV胶体金检测法和广泛应用于MVV 检测的进口商品化MVV抗体ELISA检测试剂盒对40份疑似病例血清样品进行对比检测,结果显示:用胶体金检测方法检出MVV抗体阳性血清17份,阴性血清2 3份;用进口商品化MVV抗体ELISA检测试剂盒检出阳性血清16份,阴性血清24份,二者相比整体符合率为97.5％.综上,成功建立了 MVV胶体金免疫层析快速检测方法,该方法研制的检测试纸条性能良好、便于储存、适合现场快速检测和批量检测,对MVV的监测、防控有重要意义,为内蒙古地区MVV的净化提供了新的检测方案.

外文标题：Establishment and application of colloidal gold immunochromatographic assay for detecting Maedi-Visna virus based on CA-TM fusion protein

外文摘要：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.

外文关键词：

Maedi-Visna virusprokaryotic expressionfusion proteincolloidal goldimmunochro-matography

作者：

陈思旭、包磊、李慧萍、张良、刘淑英

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作者单位：

内蒙古农业大学兽医学院,内蒙古呼和浩特 010018

农业农村部动物疾病临床诊疗技术重点实验室,内蒙古呼和浩特 010018

内蒙古自治区基础兽医学重点实验室,内蒙古呼和浩特 010018

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

基金：

国家自然科学基金项目国家自然科学基金项目内蒙古科技重大专项计划项目内蒙古草原英才创新团队项目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内蒙古自治区研究生科研创新项目

项目编号：

32072819323608632021ZD001020151031NMGIRT2412B20210174Z

出版年：

2024

DOI：

10.16656/j.issn.1673-4696.2024.0138

中国兽医科学

中国农业科学院兰州兽医研究所

中国兽医科学

CSTPCD北大核心

影响因子：0.524

ISSN：1673-4696

年,卷(期)：2024.54(8)

参考文献量11