论文部分内容阅读
目的 :研究丙型肝炎病毒 (HCV)表位抗原 ,以满足丙肝抗体确认试剂研究的需要。方法 :构建原核融合表达载体pBVIL1,精选HCV不同区抗原 :C区 ,NS3,NS4和NS5的主要抗原表位 ,用PCR方法从不同HCV抗原表达质粒中扩增出相应抗原表位 ,构建pBVIL1表达质粒 ,并在HB10 1受体菌中表达。纯化的单片段抗原经包被测定板和用间接ELISA法对阴性及阳性血清测定其活性。结果与结论 :所克隆的单片段抗原均在表达载体pBVIL1中获得高效表达 ,单片段抗原纯品用卫生部的Panel测定 ,所得结果和进口Ortho公司RIBA3.0结果进行比较 ,其中C和NS4抗原活性略高于RIBA3.0中的相应抗原 ,NS3抗原活性则略低于RIBA3.0中的c33cr抗原。但对 4 0份阳性血清的总评价和RIBA3.0完全一致 ,表明目前抗原已可满足要求
Objective: To study the antigen of hepatitis C virus (HCV) epitope in order to meet the need of confirming reagents of hepatitis C antibody. Methods: The prokaryotic expression vector pBVIL1 was constructed. The major antigenic epitopes of different regions of HCV: C region, NS3, NS4 and NS5 were selected. The corresponding antigen epitopes were amplified from different HCV antigen expression plasmids by PCR to construct pBVIL1 The plasmid was expressed and expressed in HB10 1 recipient bacteria. Purified monomeric antigens were coated on assay plates and negative and positive sera were assayed for activity using an indirect ELISA. RESULTS AND CONCLUSION: The cloned single fragment antigens were highly expressed in the expression vector pBVIL1. The single fragment antigens were purely tested by the Ministry of Health’s Panel. The results were compared with those of Ortho RIBA3.0. Among them, C and NS4 antigens Slightly higher than the corresponding antigen in RIBA3.0, and the NS3 antigen activity is slightly lower than the c33cr antigen in RIBA3.0. However, the total score of 40 positive sera was exactly the same as that of RIBA3.0, indicating that the current antigens are satisfactory