关键词：肝炎病毒组，丙型 β-半乳糖苷酶类 酶联免疫吸附测定 重组融合蛋白质类
【摘要 】 目的 为了降低检测丙型肝炎(丙肝)病毒(HCV)抗体的假阳性，建立基于双抗原夹心法的酶联免疫吸附测定(ELISA)系统。方法 利用基因工程手段，在大肠杆菌中表达HCV嵌合抗原(含核心、C33c，NS4 及NS5 抗原)与大肠杆菌β-半乳糖苷酶的融合蛋白，以该融合蛋白为酶结合物进行ELTSA。结果 构建了重组质粒，并在大肠杆菌中表达了融合蛋白，融合蛋白具有HCV抗原的抗原性，并且保持了的酶活性。以重组融合蛋白为酶结合物，用于检测抗-HCV抗体时，阳性检出率和阴性特异性均接近使用辣根过氧化物酶系统的水平。灵敏度接近以辣根过氧化物酶为标记酶的ELISA系统。结论 重组HCV抗原-β半乳糖苷酶融合蛋白可以用作酶结合物，应用于HCV感染的诊断。
HCV antibodies detection using recombinant β -galactosidase - HCV antigens chimerical protein as enzyme conjugate
CAO Cheng, LI Ping, SHI Chenghua, et al.
Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100850
【 Abstract 】 Objective To establish an ELISA system for the detection of anti-HCV antibodies by using recombinant β -galactosidase - HCV antigens chimerical prote as enzyme conjugate. Methods A four-valent HCV chimeric antigen gene was genetically fused with E.coli β -galactosidase gene (lacZ) at N terminal, and the recombinant fusion protein was used as enzyme conjugate in the ELISA system for the detection of anti-HCV antibodies.Results The gene fusion was expressed in E.coli under the control of coli phage T7 promoter and the recombinant fusion protein retained the enzyme activity of β -galactosidase as well as antigenicity of HCV antigens. It was used as enzyme conjugate in antigen sandwiched ELISA of anti-HCV antibodies using new chromogenic substrates for β -galactosidase, and similar sensitivity and specificity were obtained. Conclusion Enzyme active chimerical protein of β -galactosidase and antigens could be used as enzyme conjugate in the ELISA system when new chromatogenic substrates were used.
......
您现在查看是摘要页，全文长 13200 字符。