血清中丙型肝炎NS3抗原ELISA检测方法的建立和初步应用

The clinical application and analysis of hepatitis C virus NS3 antigen detection by ELISA in human serum

目的评价血清中丙型肝炎病毒（HCV）游离Ns，抗原的酶联免疫吸附（ELISA）检测方法的特异性和灵敏度，初步探讨该方法在临床应用中的意义。方法对77例正常人血清标本，173例抗-HCV阳性标本和3708例抗-HCV阴性的其他类型肝炎血清标本检测HCV游离NS，抗原；对部分HCVNS，抗原阳性标本进行验证，包括HCVRNA测定、中和试验和免疫斑点试验；对11例患者的25份系列血清标本进行rHCV游离Ns，抗原、HCVRNA和HCV抗体的联合检测，并结合临床资料综合分析。结果3708例抗-HCV阴性的其他类型肝炎血清标本中有48例为HCVNS，抗原阳性，其中3030例单纯乙型肝炎和445例其他类型肝炎血清标本中分别有44例和4例为HCVNS，抗原阳性；173例HCV抗体阳性标本中有42例为HCVNS，抗原阳性；77例正常人血清标本的HCVNS，抗原检测结果均为阴性；15例HCVNS，抗原阳性标本中有9例为HCVRNA阳性；23例HCVNS，抗原阳性标本的中和率和免疫斑点试验的阳性率分别为87．0％和69．6％；25份系列血清标本的检测结果显示其HCVNS，抗原的吸光度值与时间呈负相关，并有2例HCVNS，抗原阳性标本随着血清中HCVNS，抗原的吸光度值下降，其HCV抗体转阳。结论血清中HCV游离NS，抗原的ELISA检测方法有较好的特异性和敏感度，在发展中国家应用此方法进行HCV感染的早期诊断有-定的临床意义和推广价值。

Objective To evaluate the specificity and sensitivity of a novel hepatitis C virus NS3 antigen detection immunoassay and the potential application of this assay in clinical diagnosis. Methods Samples from 77 healthy subjects, 173 anti-HCV-positive patients and 3708 patients infected with other type of hepatitis were tested with the HCV NS3 antigen assay, some HCV NS3 antigen positive samples were vali- dated with HCV-RNA, neutralization and immunodot assays. Twenty-five sequential samples from 11 HCV NS3 antigen positive patients were subjected to kinetic study. Results Forty-eight （ 1.3% ） of 3708 anti- HCV negative samples were positive for HCV NS3 antigen. Among them, 44 of 3030 samples from patients only infected with HBV were HCV NS3 antigen-positive, 4 of the 445 samples from patients infected with other type hepatitis were HCV NS3 antigen-positive. In addition, 42 （24.3%） of 173 anti-HCV positive sam- ples were HCV NS3 antigen-positive and all 77 samples from healthy subjects were negative to HCV NS3 anti- gen assay. Of 15 HCV NS3 antigen-positive samples, 9 （60%） were HCV-RNA positive. The neutralization and positive percentage of immunodot assay for 23 HCV NS3 antigen-positive sera were 87.0% （20/23） and 69.6% （16/23） respectively. Of the 25 sequential samples from 11 HCV NS3 antigen positive patients, there was a negative correlation between the A values and the duration of test, and there were correlations among their HCV NS3 antigen, HCV-RNA and anti-HCV; In addition, the anti-HCV antibodies of two sera were detected while their A values of HCV NS3 antigen decreased gradually. Conclusion The HCV NS3 antigen detection assay showed perfect specificity and higher sensitivity, it will be useful in routine laboratories test in developing countries for earlier diagnosis of HCV infection.