白斑综合征病毒胶体金免疫层析试剂板的研制  

Preparation of coloidal gold immunochromatographic plate for white spot syndrome virum

作　　者：刘长军 王振国 陈青舟 夏武强 缪翁晟途 平夏婷 

机构地区：[1]象山县海洋与渔业质量技术检测中心,浙江宁波315700 [2]杭州南开日新生物技术有限公司,浙江杭州311215

出　　处：《中国生物制品学杂志》2017年第7期731-734,738,共5页Chinese Journal of Biologicals

基　　金：杭州市农业科研攻关专项(20150432B34)

摘　　要：目的研制检测对虾中白斑综合征病毒(white spot syndrome virum,WSSV)的胶体金免疫层析试剂板。方法分别制备抗WSSV单克隆抗体和多克隆抗体,并用胶体金标记单克隆抗体,将胶体金标记的抗WSSV单克隆抗体包被在金标垫上,抗WSSV多克隆抗体和羊抗鼠Ig G分别包被在硝酸纤维素膜的检测线和质控线上,组装成试剂板,通过双抗体夹心法检测样品中WSSV含量。确定试剂板检测WSSV的最低检出限,验证其特异性,并检测其与ELISA结果的符合率及稳定性。结果该试剂板对WSSV的最低检出限为1.0 mg/kg,特异性强,与ELISA实验结果相符,在室温条件下(20~25℃)可保存12个月。整个检测所需时间为8~10 min。结论该试剂板能快速有效地筛查WSSV,适合现场快速检测。Objective To prepare a coloidal gold immunochromatographic plate for white spot syndrome virum（WSSV）in shrimp. Methods Monoclonal and polyclonal antibodies against WSSV were prepared. The monoclonal antibody was labeled with colloidal gold and coated onto conjugate pad. The polyclonal antibody and goat anti-mouse Ig G were coated onto the test line and control line of nitrocellulose filter. The WSSV content in samples was determined by double antibody sandwich method. The minimum detection limit of WSSV by the plate was determined, while the specificity and stability were verified, and the coincidence rate of result to that of ELISA was evaluated. Results The minimum detection limit of WSSV by this plate was 1. 0 mg/kg. The plate showed high specificity, by which the test result was consistent with that by ELISA. The plate showed good stability after storage at room temperature（20 ~ 25 ℃） for12 months. Conclusion The plate was suitable for rapid field screening of WSSV.

关 键 词：白斑综合征病毒 双抗体夹心法 免疫层析 胶体金 

分 类 号：S917.4[农业科学—水产科学] R392-33[医药卫生—免疫学]