乙肝病毒E抗原和抗体双阳性者中病毒X及前C基因变异热点研究

The mutations of HBV X and precore gene isolated from both HBeAg and HBeAb positive blood donors

目的 了解乙型肝炎患者 HBeAg和 HBeAb双阳性状态下X区及前C区基因热点变异情况,探讨T1762与 A1764及 A1896热点变异与 HBe转换时相的关系。方法 采用时间分辨荧光免疫分析方法定量检测乙肝5项标志物.对HBeAg/HBeAb双阳性标本采用巢式聚合酶链反应扩增,其包括X区及前C区在内的DNA片段,并对阳性的PCR产物直接标记测序,测序结果和Genbank中登录的标准序列相比较。结果 对15例HBeAg和HBeAb双阳性患者血清中HBV DNA进行了检测,阳性11例,测序结果显示,11例HBV DNA阳性者均存在T1762和A1764的突变,但仅有4例患者出现了AI896的突变。结论 在乙型肝炎HBe转换过程中均伴有BCP区T1762和A1764的突变,部分存在A1896位点的突变,T1762和 A1764的突变要早于 A1896的突变,而A1896的突变主要在E抗体产生过程中或产生以后。

Objective To investigate the mutations of HBV X and precore gene isolated from both HBeAg and HBeAb positive blood donors. Methods HBV markers were detected by DELFIA, HBV X and precore gene were amplified by polymerase chain reaction and sequenced directedly in ABI 310 machine. The X and precore gene sequences were compared with the representative sequences in Genbank, then the mutations were found. Results Eleven of 15 patients were positive for HBV DNA. The sequence analysis showed that mutations existed all sequences. T1762 and A1764 mutations were found in all patients and A1896 mutations were found in four patients. Conclusion There are X and precore gene mutations including those of T1762, T1764 and part of A1896 in patients with HBe seroconversion. T1762 and A1764 are detected before A1896, and A1896 mutations may be detected during or after production of HBeAb in patients with hepatitis B virus infection.