【摘要】 目的：在大肠杆菌中表达丙型肝炎病毒（ｈｅｐａｔｉｔｉｓ Ｃｖｉｒｕｓ，ＨＣＶ）Ｅ２ 包膜蛋白，并对其进行初步纯化。用纯化的包膜蛋白检测ＨＣＶＲＮＡ 阳性血清中Ｅ２ 抗体，研究其检测Ｅ２ 抗体的敏感性和特异性。方法：将ＨＣＶＥ２ 蛋白基因克隆到原核表达载体中，并转化大肠杆菌。用ＩＰＴＧ 诱导细菌表达Ｅ２ 蛋白，经离子交换层析纯化蛋白。用纯化的包膜蛋白包被微孔板，经ＥＬＩＳＡ方法检测ＨＣＶＲＮＡ阳性血清中的Ｅ２ 抗体。结果：在大肠杆菌表达了相对分子质量为４５ ０００ 的可溶性Ｅ２ 蛋白，占细菌总蛋白量的２１％ ，纯化后，蛋白纯度为８５％ 。用其检测ＨＣＶ 患者血清１７１ 例，Ｅ２ 抗体的检出率为６３％ 。在Ｅ２ 抗体阳性的血清中，部分为抗ＨＣＶ 阴性。结论：Ｅ２ 蛋白的表达及初步纯化的成功，可能用于临床检测。各基因型ＨＣＶＥ２ 包膜蛋白的抗体之间有交叉反应，在目前的抗ＨＣＶ 诊断试剂中加入可溶性、非融合Ｅ２ 包膜蛋白，可以改善抗ＨＣＶ 诊断试剂的特异性和敏感性。更多还原

【Abstract】 Objective: To express HCV E2 protein in E.coli and study the specificity and sensitivity of anti E2 antibody measurement. Methods: The vector expressing HCV E2 protein was constructed by inserting E2 gene into prokaryotic expression vector and transformed into E.coli , then the E.coli was induced with IPTG to express E2 protein. By means of affinity chromatography the expressed protein was purified. By ELISA, the anti E2 antibodies were detected in HCV RNA positive serum using the expressed protein. Results: The soluble E2 protein with the molecular mass of 45 000 was expressed in E.coli . After purification, the purity of the E2 protein was 85 percent. The purified E2 protein used, anti E2 antibodies can be detected in 63% patients with chronic hepatitis C and can be detected in some HCV RNA positive but anti HCV negative serum. Conclusion: The success of expressing and purifying HCV E2 protein was helpful for studying the clinical significance of anti E2 antibodies examination. The antibodies against E2 from a HCV isolate can crossly react with the E2 protein from another isolate. Inclusion of E2 protein in the antigenic mixture utilized in HCV screening assay would improve the sensitivity and specificity of serological assays.更多还原