丙型肝炎病毒第一高变区嵌合抗原的纯化及其在血液筛查中的应用

发布时间：2018-11-20 08:01

【摘要】： 丙型肝炎病毒(HCV)属黄热病毒科,感染人体后可引起机体免疫应答发生异常,导致组织损伤,形成慢性持续性感染,并能进一步发展为肝硬化甚至肝细胞肝癌。HCV整个基因组呈高度变异,其中位于HCV包膜蛋白E2 N端的27个氨基酸变异最大,故命名为第一高变区(HVR1)。丙型肝炎病毒抗体检测试剂在血液筛查、临床疾病诊断中发挥着重要作用。但由于HCV的高变异,致使现有HCV抗体检测试剂存在漏检、中和性抗体难以检测等问题。军事医学科学院基础医学研究所利用生物信息学筛选出4条具有高交叉反应性的HVR1抗原,并利用自行构建的pBV IL1载体进行了上述HVR1抗原的嵌合表达,获得了1条能覆盖大部分HVR1抗原变异的嵌合抗原F4HVR1 (专利号: ZL 0114187916)。本研究在此基础上,对嵌合抗原进行纯化,获得蛋白纯品,鉴定嵌合抗原的活性和特异性,建立了HVR1抗体ELISA检测技术。本研究对北京市红十字血液中心2008-2009年间的77991名献血者的ALT和乙肝表面抗原(HBsAg)检测结果进行了统计和分析,研究ALT不合格者的性别、年龄、季节分布特点,并作出了ALT不合格与HBsAg的相关性分析。本研究对上述ALT单项不合格中的2035份血清进行了HVR1抗体的检测,并与C区、NS3区、NS4区、NS5区抗体的检测作比较,确立HVR1单独抗体阳性血清,评价HVR1抗体检测在防止HCV血液漏检中的重要作用;建立了用于评价HVR1抗体检测试剂的献血者血清质控品。该抗原与目前HCV诊断试剂中采用的保守抗原完全不同,能从本质上提高HCV血液筛查的灵敏度。
[Abstract]:Hepatitis C virus (HCV) belongs to the yellow fever virus family. It can cause abnormal immune response, tissue damage and chronic persistent infection. The whole genome of HCV was highly variable, of which 27 amino acids located at the N terminal of HCV envelope protein E2 had the largest variation, so it was named the first hypervariable region (HVR1). Hepatitis C virus antibody detection reagent plays an important role in blood screening and clinical diagnosis. However, due to the high variation of HCV, the existing HCV antibody detection reagent has some problems, such as missed detection and difficult detection of neutralizing antibody. The Institute of basic Medicine, Academy of military Medical Sciences, screened out four HVR1 antigens with high cross-reactivity by bioinformatics, and expressed the HVR1 antigen by using the self-constructed pBV IL1 vector. A chimeric antigen F4HVR1 (patent number: ZL 0114187916) covering most of the variation of HVR1 antigen was obtained. In this study, the chimeric antigen was purified, protein purity was obtained, the activity and specificity of chimeric antigen were identified, and the detection technique of HVR1 antibody ELISA was established. In this study, the results of ALT and hepatitis B surface antigen (HBsAg) detection in 77991 blood donors from Beijing Red Cross Blood Center in 20082009 were statistically analyzed, and the sex, age, seasonal distribution characteristics of ALT unqualified patients were studied. The correlation between ALT disqualification and HBsAg was analyzed. In this study, 2035 unqualified ALT sera were tested for HVR1 antibody, and compared with C region, NS3 region, NS4 region and NS5 region. To evaluate the important role of HVR1 antibody detection in preventing HCV blood leakage. The serum quality control products of blood donors for the evaluation of HVR1 antibody detection reagent were established. The antigens are completely different from the conservative antigens used in HCV diagnostic reagents and can improve the sensitivity of HCV blood screening essentially.
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R392.1

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