牛传染性鼻气管炎病毒胶体金检测试纸条的研制和评价  

Development and Evaluation of Colloidal Gold Test Strip for Detection of Infections Bovine Rhinotracheitis Virus

作　　者：刘文晓 张坤 程晶 李永清[1] LIU Wenxiao;ZHANG Kun;CHENG Jing;LI Yongqing(Animal Disease Research Center,Beijing Academy of Agriculture and Forestry Sciences,Beijing 100089,China;Animal Science and Technology College,Beijing University of Agriculture,Beijing 100096,China)

机构地区：[1]北京市农林科学院畜禽疫病研究中心,北京海淀100089 [2]北京农学院动物科学技术学院,北京昌平100096

出　　处：《中国兽医杂志》2024年第11期70-76,共7页Chinese Journal of Veterinary Medicine

基　　金：国家自然科学基金面上项目(32172818);北京市自然科学基金面上项目(6232012);北京市农林科学院科技创新能力建设专项(KJCX20200424);家畜产业技术体系北京市创新团队项目(BAIC06-2022)。

摘　　要：为建立快速检测牛传染性鼻气管炎病毒(IBRV)的病原学检测方法,本试验采用杂交瘤细胞融合技术制备IBRV单克隆抗体,以其作为检测原,优化反应条件并组装IBRV胶体金检测试纸条,验证该试纸条的特异性、灵敏性、敏感性和稳定性,并与实时荧光定量PCR(qPCR)进行比较分析。结果显示,共制备7株针对IBRV的单克隆抗体,其中以胶体金标记的单克隆抗体2B7作为捕获抗体、1B9作为检测抗体,建立的胶体金检测试纸条能够检测样品中的IBRV;该试纸条与病毒性腹泻/黏膜病毒、牛冠状病毒和大肠杆菌无交叉反应,检测限为4.04×10~4TCID50/0.1 mL,敏感性为86.7%;在室温条件下保存12个月仍能够有效检测IBRV,说明该试纸条稳定性良好。与qPCR比较结果显示,2种方法检测结果的符合率为92.85%,说明该胶体金检测试纸条能够用于IBRV的临床诊断。结果表明,本试验利用IBRV的单克隆抗体建立的胶体金检测试纸条灵敏度高且特异性好,为IBRV抗原检测和流行病学调查提供技术支持。To develop a rapid pathogen detection method for infectious bovine rhinotracheitis virus(IBRV),this study utilized hybridoma technology to prepare monoclonal antibodies(mAbs)against IBRV.These mAbs were used to optimize the reaction conditions and assemble a colloidal gold test strip for IBRV detection.The test strip's specificity,sensitivity,and stability were validated and compared with real-time quantitative polymerase chain reaction(qPCR).The results showed that seven monoclonal antibodies targeting IBRV were produced.Using mAb 2B7 labeled with colloidal gold as the capture antibody and mAb 1B9 as the detection antibody,the developed colloidal gold test strip could detect IBRV in samples.The test strip showed no cross-reactivity with bovine viral diarrhea/mucosal virus,bovine coronavirus,or Escherichia coli,with a detection limit of 4.04×104 TCID 50/0.1 mL and a sensitivity of 86.7%.The test strip remained effective for IBRV detection after 12 months of storage at room temperature,indicating good stability.When compared to qPCR,the two methods had a concordance rate of 92.85%,demonstrating that the colloidal gold test strip is suitable for clinical diagnosis of IBRV.These results suggest that the colloidal gold test strip developed using IBRV monoclonal antibodies is highly sensitive and specific,providing technical support for IBRV antigen detection and epidemiological surveys.

关 键 词：牛传染性鼻气管炎病毒(IBRV) 胶体金检测试纸条 单克隆抗体 

分 类 号：S855.3[农业科学—临床兽医学]