胶体金渗滤方法检测乙肝患者血清中乙肝核心抗体的研究  

Detection of Ig M anti-Hbs Ag in Serum by Immnogold Filtration

作　　者：张爱英[1] 杨国申 李康[3] 赵元顺[3] 陈怡[4] 侯传浩 李宁[1,3] 

机构地区：[1]首都医科大学附属北京佑安医院北京市肝病研究所,北京100069 [2]首都医科大学燕京医学院,北京100080 [3]首都医科大学附属北京佑安医院,北京100069 [4]首都医科大学基础医学院,北京100069

出　　处：《现代生物医学进展》2014年第36期7165-7167,7059,共4页Progress in Modern Biomedicine

基　　金：北京市肝病研究所基金(BJIH-01206);北京佑安医院肝病艾滋病基金;首都医科大学重点实验室基金(2013GYGA03);国家国际科技合作专项<中美英联合乙肝相关肝癌早期诊断分子标志物筛查及免疫机制研究>(S2012ZR0153)

摘　　要：目的:建立胶体金免疫渗滤法检测乙肝患者血清中乙肝核心抗体(anti-HBc Ig M)抗体的方法,评价其灵敏度、特异性和重复性。方法:预先在硝酸纤维素膜上包被乙肝核心抗原HBc Ag,将血清或者血浆加在硝酸纤维素膜上,血清或者血浆中的anti-HBc Ig M与HBc Ag结合。洗涤去掉非特异结合,加入胶体金标记的抗人-Ig M,洗涤去掉没有结合的抗人Ig M抗体。硝酸纤维素膜上的抗原-Ig M-抗人Ig M形成的"三明治"复合物呈现红色圆点,证实实验有效。结果:102份乙肝患者血清,anti-HBc Ig M阳性标本99份。检出率97%。50份正常血清中,有一份检测结果弱阳性,特异性98%。结论:胶体金免疫渗滤法检测anti-HBc Ig M敏感性高、特异性强、重复性好,且方便快捷,不需要特殊的仪器设备,在anti-HBc Ig M快速检测上有广阔的应用前景。Objective： To develop the immunogold filtration assay（IGFA）method for the detection of anti-HBc Ag Ig M and evaluate its sensitivity, specificity and reproducibility. Methods： Nitrocellulose membranes were coated with Hbc Ag. Patient＇s serum or plasma sample is added to the Nitrocellulose membranes, and the anti-HBc Ig M was bound to the pre-coated HBc Ag. After washed to remove sample serum proteins, rabbit anti-Human Ig M labeled with colloidal gold was added. After washing to remove unbound anti-Human Ig M, in presence of the antigen-Ig M- anti-Human Ig M ＂sandwich＂ complex in a red dot, which indicates the validity of the test.Results： The results showed that 99 out of 102 total cases were positive in serum of patients with hapetitis. The sensitivity of the IGFA was 97%. The specificity was 98% in 50 healthy individuals（1/50）. Conclusion： IGFA has high sensitivity, strong specificity and good reproducibility and the advantage of convenient and without equipment or the detection of Hbc Ab. It is potential rapid tool for the detection of Hbc Ab in serum samples.

关 键 词：胶体金免疫渗滤 乙肝核心抗体 IG M 

分 类 号：R512.62[医药卫生—内科学] R446[医药卫生—临床医学]