用酶联免疫夹心法测定肝炎患者血清丙型肝炎抗原  

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

作　　者：谢立[1] 黄德庄[1] 时洪波[1] 贺丽香[1] 靳海英[1] 

机构地区：[1]首都医科大学附属北京佑安医院,北京市卫生局肝炎研究所100054

出　　处：《中华检验医学杂志》2005年第11期1159-1162,共4页Chinese Journal of Laboratory Medicine

基　　金：首都医学发展科研基金课题(2002-3068);首都医科大学基础临床合作基金课题(02JL20)

摘　　要：目的建立血清中丙肝病毒NS3抗原(HCAgNS3)的酶免检测方法。方法以特异性强、敏感度高、识别不同HCAgNS3抗原表位的HCAgNS3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HCAgNS3抗原。结果优化了酶免疫反应条件;该方法对HCAgNS3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HCAgNS3抗原的检出率为21.2%,对15例HCAgNS3阳性标本测定HCVRNA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,HCAgNS3检出率为1.7%,50例正常人血清标本的HCAgNS3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。Objective To establish the enzyme linked immunosorbent assay （ELISA） for detecting the Hepatitis C virus NS3 antigen from serum samples. Methods A sandwich ELISA for detecting the Hepatitis C virus NS3 antigen was developed by using perfect specificity and higher sensitivity anti-HCV NS3 Ag McAb and high efficient purified anti-HCV polyclone antibody. Results The test could detect HCV NS3 antigen of 1 - 2ng/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 antiHCV were positive to HCV NS3 antigen. Nine of 15 samples, which were positive to HCV NS3 antigen, were positive to HCV RNA. Fifty normal serum samples were negative to HCV NS3 antigen. Conclusion The HCV NS3 antigen detection method showed perfect specificity, high sensitivity and well stabilization. It can be used for the early diagnosis of HCV.

关 键 词：C型肝炎样病毒属 肝炎抗原 丙型 抗体 单克隆 酶联免疫吸附测定 

分 类 号：R446.6[医药卫生—诊断学]