胶体金免疫斑点法检测人巨细胞病毒pp150抗体

Detection of IgM antibodies to HCMV pp150 by colloidal gold immunodotting assay

目的 建立一种简便、快速的检测血清中人巨细胞病毒 (HCMV)抗 pp15 0抗体的胶体金免疫斑点法。 方法 采用Frens法制备胶体金颗粒 ,标记葡萄球菌A蛋白 (SPA) ,将本所利用基因工程技术生产的 pp15 0抗原固定于 0 45 μm孔径的硝酸纤维素膜上 ,制成免疫斑点检测装置。血清中抗pp15 0IgG与硝酸纤维素膜上的抗原结合 ,然后滴加胶体金标记的SPA ,在膜上可形成肉眼可见的红色斑点。结果 胶体金免疫斑点法与ELISA试剂盒对比检测了 2 0 0份临床确诊血清标本中抗 pp15 0IgG抗体 ,胶体金法的特异性和敏感性分别为 92 %和 90 %。 结论 胶体金免疫斑点法检测血清中的抗 pp15 0抗体特异性、灵敏度较高 ,简便快速 。

Objective To develop a rapid and simple method to detect the human IgM antibodies to cytomegalovirus (HCMV) pp150 in sera by colloidal gold immunodotting assay.Methods HCMV pp150 protein was produced by gene engineering.Colloidal gold immunodotting apparatus was prepared with nitrocellulose filter of 0.45 μm poresize.The colloidal gold particles with 10 nm diameter were prepared by Frens method and were labeled with Staphylococcal protein A (SPA).The gold-SPA conjugates reacted with human pp150 IgM antibodies in sera and then a visible red dot on the nitrocellulose which the carboxyl terminus of pp150 protein were immobilized on was formed.Results The pp150 IgM antibodies in 200 sera were detected by both colloidal gold immunodotting assay and enzyme immunoassay (EIA) simultaneously.The specificity and sensitivity of the former were 95% and 90% respectively.Conclusion The colloidal gold immunodotting assay is a sensitive,specific,simple and rapid assay for detecting pp150 IgM antibodies in sera.It is high promising for broad applications.