张爱英1# 杨国申2# 李康3 赵元顺3 陈怡4 侯传浩4 李宁1,3△.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[J].现代生物医学进展英文版,2014,14(36):7165-7167. |
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 IgManti-HbsAg in Serumby Immnogold Filtration |
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DOI: |
中文关键词: 胶体金免疫渗滤 乙肝核心抗体 IgM |
英文关键词: Immunogold filtration assay HBcAb IgM |
基金项目: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) |
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中文摘要: |
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 IgM)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HBcAg,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
IgM与HBcAg结合。洗涤去掉非特异结合,加入胶体金标记的抗人-IgM, 洗涤去掉没有结合的抗人IgM抗体。硝酸纤维素膜上的
抗原-IgM-抗人IgM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 IgM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 IgM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 IgM快速检测上有广阔的应用前景。 |
英文摘要: |
Objective:To develop the immunogold filtration assay (IGFA)method for the detection of anti-HBcAg IgM and
evaluate its sensitivity, specificity and reproducibility.Methods:Nitrocellulose membranes were coated with HbcAg. Patient's serum or
plasma sample is added to the Nitrocellulose membranes, and the anti-HBc IgM was bound to the pre-coated HBcAg. After washed to
remove sample serum proteins, rabbit anti-Human IgMlabeled with colloidal gold was added. After washing to remove unbound anti-
Human IgM, in presence of the antigen-IgM- anti-Human IgM "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 HbcAb. It is potential rapid tool for the
detection of HbcAb in serumsamples. |
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