血清丙型肝炎病毒抗原的免疫酶斑点法检测

目的建立一种可筛查早期HCV感染者的简单快捷方法。方法制备特异性的抗HCV核心蛋白和抗HCVNS3的单克隆抗体(mAb)酶标记物,应用免疫酶斑点法检测各种类型肝炎患者和正常人血清中的HCV-Ag,并与双抗体夹心ELISA检测HCV-Ag及RT-PCR检测HCVRNA的结果进行比较。结果免疫酶斑点法检测表明,HCV-Ag的检出率在抗HCV抗体阳性、乙肝和非甲非戊肝炎患者中,分别为15.4%(8/52)、1.73%(19/1099)和1.03%(1/97);在81例甲肝和67例戊肝患者及50例正常人中均未检出。免疫酶斑点法与双抗体夹心ELISA及RT-PCR的检测的结果没有显著性差异,但其与RT-PCR检测结果有一定的相关性。结论免疫酶斑点法特异性强,操作简便快捷,不需要特殊仪器设备,为HCV的早期诊断和常规筛查提供了有效的方法。

Detection of HCV antigen in the serum by immunodotting

AIM: To set up a simple and rapid screening method of early dection for HCV infection. METHODS: The HRP-anti-HCV monoclonal antibodies (mAbs) conjugate was prepared by the mAbs of anti-HCV-core and anti-HCV-NS_3 with high specificity. HCV antigens in the sera from hepatitis patients and normal persons were detected by immunodotting and compared by double antibody ELISA and RT-PCR. RESULTS: The positive ratio for HCV antigen was 15.4%(8/52), 1.73%(19/1 099) and 1.03%(1/97) in hepatitis C patients and hepatitis B patients and hepatitis NA-NE patients, respectively. The results for HCV antigen were negative in hepatitis A patients (0/81), hepatitis E patients (0/67) and normal persons(0/50). There was no significant difference among the immunodotting and double antibody ELISA and RT-PCR. There was definite correlation between the immunodotting and RT-PCR. CONCLUSION: This method, simple and rapid with high specificity, provides an effective way for the early diagnosis and routine detection of HCV.