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HBVX蛋白(HBx)是由乙型肝炎病毒(HBV)基因组编码的调控蛋白,与由乙肝引起的肝癌发生具有密切的关系。血清HBx对乙型肝炎、肝癌的诊断及发病机理研究有较高的临床应用价值。在表达并纯化HBx、制备出HBx单克隆抗体与酶标抗体的基础上,研制了HBx定量检测试剂盒(增强化学发光法),对其灵敏度、特异性等指标进行分析,并将该试剂盒应用于临床研究。结果表明,原核表达并纯化的HBx纯度≥94%;试剂盒灵敏度达0.1ng/ml;线性范围达到0.5～600ng/ml;特异性高,与球蛋白、脂蛋白、血红蛋白、酸性糖蛋白、HBc、HBe、HBs、HBV preS2不发生交叉反应;批间CV≤6.5%;临床标本检验慢性乙型肝炎阳性率为55%、肝硬化阳性率为68%、肝癌阳性率70%,表明此试剂盒可应用于乙肝、肝硬化、肝癌的临床诊断及发病机理研究。
HBVx protein (HBx) is a regulatory protein encoded by the hepatitis B virus (HBV) genome and has a close relationship with the occurrence of hepatocellular carcinoma caused by hepatitis B virus. Serum HBx on hepatitis B, liver cancer diagnosis and pathogenesis of high clinical value. Based on the expression and purification of HBx and the preparation of HBx monoclonal antibody and enzyme-labeled antibody, the HBx quantitative detection kit (enhanced chemiluminescence method) was developed and its sensitivity, specificity and other indicators were analyzed. The kit Used in clinical research. The results showed that the purity of prokaryotic expression and purification of HBx was ≥94%, the sensitivity of the kit was 0.1ng / ml, the linear range was 0.5-600ng / ml, the specificity was high, and the specificity of globulin, lipoprotein, hemoglobin, acid glycoprotein, HBc , HBe, HBs, HBV preS2 do not cross-react; Inter-CV = 6.5%; clinical specimens of chronic hepatitis B positive rate of 55%, cirrhosis positive rate of 68%, liver cancer positive rate of 70%, indicating that the kit Can be applied to hepatitis B, cirrhosis, liver cancer clinical diagnosis and pathogenesis.