以抗原半乳糖苷酶融合蛋白为酶结合物检测丙型肝炎抗体  

HCV antibodies detection using recombinant β galactosidase HCV antigens chimerical protein as enzyme conjugate

作　　者：曹诚[1] 李平[1] 石成华[1] 马清钧[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100850

出　　处：《中华医学检验杂志》1999年第6期333-335,共3页

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。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.

关 键 词：丙型肝炎 抗体 ELISA HCV Β-半乳糖苷酶 

分 类 号：R512.630.4[医药卫生—内科学] R446.62[医药卫生—临床医学]