庚型肝炎病毒NS3区抗原基因的原核表达

Prokaryotic expression of antigen gene within HGV NS3 region and preliminary application

目的 建立检测HGV抗体酶联免疫吸附试验 (ELISA)的方法。方法 选取HGVNS3区具有较强免疫原性的C17抗原基因 ,构建原核表达质粒 ,在大肠杆菌中进行诱导表达。结果 C17抗原基因获得了正确而高效的表达 ,经纯化、复性后建立的ELISA技术与RT PCR法同时检测血清标本 ,其灵敏度和特异度分别为 40 %和 98.4%,检测结果一致率为 82 .2 %。联合包被另外两种非结构区蛋白 (NS4,NS5 ) ,可使灵敏度提高到 6 0 %,而保持一定的特异度。结论 该方法检测HGV抗体可靠 ,可作为临床应用。

Objective To set up a ELISA method to test hepatitis G virus(HGV) antibody. Methods The selected C17 gene was believed a strong immunogen-coding sequence within HGV NS3 region and constructed a prokaryotic expression plasmid named pQEHGVCl7 gene. The target gene was expressed accurately and effectively in E.coli. Then antigen C17 protein was obtained via purification and used asanagent for HGV antibody test. Results It was found that the sensitivity and specificity of ELISA were 40% and 98.4%, respectively. If two more antigen proteins (within NS3. NS4. NS5) have been coated simultaneously, the sensitivity could raise to 60% while the specificity keep the same level. Conclusion The method is an effective method and can be used in clinical study.