用酶联免疫夹心法测定肝炎患者血清丙型肝炎抗原

目的建立血清中丙肝病毒NS3抗原（HCAg-NS3）的酶免检测方法。方法以特异性强、敏感度高、识别不同HCAg-NS3抗原表位的HCAg-NS3单克隆抗体和高效价的纯化抗-HCV分别作为包被抗体和酶标记抗体，采用夹心ELISA法测定血清中HCAg-NS3抗原。结果优化了酶免疫反应条件；该方法对HCAg-NS3的最低检测限为1—2ng／ml；批内（n=13）及批间（n=3）变异系数（CV）分别为4．51％和5．64％；对52例抗-HCV阳性血清测定HCAg-NS3抗原的检出率为21．2％，对15例HCAg-NS3阳性标本测定HCV RNA的检出率为60％，1350例抗-HCV阴性的其他类型肝炎患者血清标本中，HCAg-NS3检出率为1．7％，50例正常人血清标本的HCAg-NS3抗原测定结果均为阴性。结论该方法敏感性较高，特异性、重复性和稳定性均良好，可以作为早期诊断HCV感染的有效方法。

The detection of Hepatitis C virus NS3 antigen in human serum by sandwich enzyme-linked immunoassay

Objective To establish the enzyme linked immunosorbent assay (ELISA) for detecting the Hepatitis C virus NS_3 antigen from serum samples.Methods A sandwich ELISA for detecting the Hepatitis C virus NS_3 antigen was developed by using perfect specificity and higher sensitivity anti-HCV NS_3 Ag McAb and high efficient purified anti-HCV polyclone antibody.Results The test could detect HCV NS_3 antigen of 1~2 ng/ml.The CVs of intra-group (n=13) and inter-group (n=3) were 4.51% and 5.64% respectively.Eleven of 52 samples being positive to anti-HCV and 23 of 1350 ones being negative to anti-HCV were positive to HCV NS_3 antigen.Nine of 15 samples, which were positive to HCV NS_3 antigen, were positive to HCV RNA. Fifty normal serum samples were negative to HCV NS_3 antigen.Conclusion The HCV NS_3 antigen detection method showed perfect specificity, high sensitivity and well stabilization. It can be used for the early diagnosis of HCV.