猪伪狂犬病毒gB抗原侧向层析快速检测试纸条的研制  

Preparation of a Lateral Flow Assay for Detection of Glycoprotein B of Pseudorabies Virus in Swine Herds

作　　者：王华俊[1] 班付国[1] 赵雪丽[1] 谢彩华[1] 闫若潜[1] 王淑娟[1] 马震原[1] 王东方[1] 王翠[1] Wang Huajun;Ban Fuguo;Zhao Xueli;Xie Caihua;Yan Ruoqian;Wang Shujuan;Ma Zhenyuan;Wang Dongfang;Wang Cui(Henan Animal Diseases Prevention and Control Centre,Zhengzhou,Henan 450008,China)

机构地区：[1]河南省动物疫病预防控制中心,河南郑州450008

出　　处：《中国动物检疫》2018年第12期85-90,共6页China Animal Health Inspection

基　　金：河南省科技创新人才计划项目(174200510003)

摘　　要：为建立一种简便、快速、特异的猪伪狂犬病毒(PRV)gB抗原检测方法,利用单克隆抗体技术和侧向层析(LFA)技术,采用柠檬酸三钠还原法,制备颗粒大小为31.5 nm胶体金,再用胶体金标记纯化的抗PRV gB单克隆抗体10D4,在硝酸纤维素膜的质控带和检测带处,分别包被羊抗鼠IgG二抗和单克隆抗体6E9,组装成LFA方法通用胶体金层析试纸条。经测试,该试纸条最低能检测出1:640倍稀释的,TCID_(50)为1×10^(8.6)/0.1 mL的灭活PRV抗原,并与古典猪瘟病毒(CSFV)、猪繁殖与呼吸综合征病毒(PRRSV)、猪圆环病毒2型(PCV2)、猪细小病毒(PPV)、猪流行性腹泻病毒(PEDV)均无交叉反应,特异性良好;室温干燥密闭的条件下,该试纸条至少可保存10个月。结果表明,本研究建立的试纸条方法操作简单、快速、敏感、特异,易于判定,适合基层PRV的大面积普查和现场检测。In order to establish a simple,rapid and specific method for detection of pseudorabies virus(PRV)gB antigen in swine herds,the monoclonal antibody technology and lateral flow assay(LFA)were applied.Firstly,the colloidal gold particles of31.5nm were synthesized by trisodium citrate reduction method,then the purified monoclonal antibody of10D4was labeled with the synthesized colloidal gold particles.The goat anti-mouse IgG secondary antibody and monoclonal antibody of6E9were coated in the quality-control zone and detection zone of the nitrocellulose membrane,respectively.At last,the LFA test strips were assembled.The results showed that the strips could detect inactivated PRV antigens which were made a640fold dilution(TCID50:1×10^8.6/0.1mL).Besides,the method had a good specificity,hence no cross-reaction with CSFV,PRRSV,PCV2,PPV and PEDV was found.Under the conditions of room temperature,desiccation and being sealed,the strips could be stored for at least10months.As a conclusion,the lateral flow assay established in this study was simple,rapid,sensitive,specific and convenient.It was suitable for PRV large-area census and on-site detection at the grass-roots.

关 键 词：猪 伪狂犬病毒 侧向层析 单克隆抗体 试纸条 

分 类 号：S852.65[农业科学—基础兽医学]