目的 建立检测抗-HCV的斑点酶免疫法。方法 将基因重组HCV核心、NS3/NS4、NS5抗原和马抗人IgG、人IgG以最佳浓度固定在硝酸纤维膜条带上，用辣根过氧化酶和改进的底物进行斑点酶免疫反应。结果 检测了14份经ORTHO第三代抗-HCVELISA检测的血清，符合率为100%；72份经UBI第三代抗-HCVELISA监测的血清，符合率95.83%；103份经国产厦门新创抗-HCVELISA监测的血清，符合率95.14%。54份自然人群血清，阳性率3.47%。89份阳性标本中，三种抗体同时阳性的占30.34%，两种抗体阳性占69.66%，没有检出单一抗体阳性标本。核心抗体检出率最高87.64%，NS3/NS4抗体次之76.40%，NS5抗体最低61.80%。结论 用基因重组抗原建立的斑点酶免疫法与国内外广泛应用的ELISA试剂有较高的符合率，对进一步研究HCV感染诊断的确认试剂有意义。%Objective To set up a configuration of immunoblo tting assay forcletecting serum antibodies against HCV.Methods The proteins of the said virus that Core and NS3/NS4 and NS5 expres sed in E.coli cells were immobilized separately on nitrocellulose filter as antigens.HRP and a modified preparation of chromogenic substrate were used in the following procedure.Results Among 189 samples of sera tested by this Blotting Assay,14 were tested also by using ORTHO 3rd Generation ELISA regent and the coincident rate was 100% between the two assays,72 were tested by UBI 3rd Generation ELISA and their coincident rate was 95.83%，103 were tested by Xia Meng Xin Chuang ELISA and the coincident rate was 95.14%,Among 89 posi tive sera determined by Blotting Assay,those showed positive for all three antig ens accounted for 30.34%，and those positive for two of three antigens accounted for 69.66%，positive for only one antigen was not found.Among the 3 antibodies of Core was most efficient that 78 out of 89 positive sera reacted to it,the next was NS3/NS4with that number of 68/89,then SN5 with that of 55/89.Conclusion The immunoblotting assay showed comparatively high coincident with ELISA suggested its potential value in diagnosing HCV infection .
展开▼
