GLT25D1血清水平与慢性乙型肝炎肝纤维化进程相关性分析

摘要

目的 探讨GLT25D1血清水平与慢性乙型肝炎肝纤维化进程的相关性.方法 通过抗原表位预测软件BepiPred 1.0Server将GLT25D1基因片段分为GLT25D 1-1和GLT25D 1-2两部分,PCR扩增获得目的片段,构建其原核表达质粒；对制备的多克隆抗体进行效价及特异性分析.通过以GLT25D1为抗原的ELISA双抗体夹心的方法,检测其在HBV感染不同阶段的患者的血清水平.结果 成功表达重组蛋白,Western blot鉴定正确；成功制备兔抗人(GLT25D1-1、GLT25D 1-2)的多克隆抗体,效价较高,特异性良好.通过ELISA方法检测HBV感染患者血清,慢性乙肝(病理结果不超过S3期)、肝硬化(S3-4期及S4期)以及肝癌患者中GLT25D1的OD值较正常组均有显著性差异(P＜0.01).结论 通过血清学检测结果,我们发现GTL25D1存在于HBV感染的肝病的各进程中,进而我们推测血清GLT25D1可能会作为检测肝纤维化进程的一个新的血清学指标.%Objective To discuss the relationship between serum concentration of glycosyltransferase 25 domain 1 ( GLT25D1) and the grading of fibrosis in patients with chronic HBV infection. Methods The GLT25D1 gene was divided into two parts of GLT25D1-1 and GLT25D1-2 by antigenic epitope prediction software BepiPred 1.0 Server. With reverse transcription polymerase chain reaction (RT-PCR) , genes (GLT25D1-1 and GLT25D1-2) were cloned into pro-karyotic expression vector pET-32a( + ). The potency and specificity of polyclonal antibody were evaluated by enzyme-linked immunosorbent assay ( ELISA) and Western blot. An antigen to GLT25D1 ELISA double antibody sandwich method was used to detect, the serum of GLT25D1 in patients with various stages in the process of HBV infection of the liver. Results ELISA analysis indicated that the titer of the polyclonal antibody can be achieved 1: 1 280 000. The high specificity of the polyclonal antibody was confirmed by Western blot. To detect the serum of HBV infection at different stages by ELISA, we found that there were statistically significant differences of GLT25D1 OD values in chronic hepatitis B ( ≤S3) , cirrhosis (S3-4 & S4) and liver cancer patients compared with the normal group (P < 0. 01 ). Conclusion With the serological testing results of chronic hepatitis, cirrhosis, and liver cancer patients, we found GTL25D1 exist in the process of HBV infection, and thus we speculated serum GLT25D1 detection can be used as an objective indicator to detect liver injury, and to assist other detection indicators better to provide the impetus for the diagnosis of hepatitis HBV infection process.