慢性HBVC基因启动子变异的检测  

Detection of core promoter mutations in chronic hepatitis B

作　　者：邢利和[1] 朱传琳[2] 施红[2] 韩凤连[2] 

机构地区：[1]解放军第251医院,张家口075000 [2]解放军第302医院

出　　处：《中华实验和临床病毒学杂志》2000年第2期163-165,共3页Chinese Journal of Experimental and Clinical Virology

摘　　要：目的　了解慢性乙型肝炎病毒 (HBV)C基因启动子变异的情况以及对病毒血清学的影响。方法　应用错配聚合酶链反应特异性扩增含有HBVC基因启动子的片段 ,扩增产物用限制性内切酶BcLⅠ酶切 ,琼脂糖凝胶电泳后 ,观察酶切后的限制性片段长度多态性 (RFLP)图谱 ,分析C基因启动子区段的核苷酸 (nt) 176 2由碱基A→T和 (nt) 176 4碱基由G→A的变异 ,并经直接测序分析证实RFLP结果。结果　 110例慢性乙型肝炎病人中HBVDNA阳性 89例 ,其中HBeAg阳性组 43例 ,检测出T 176 2 /A 176 4变异株 2 1例 ;46例抗 HBe阳性组中检出T 176 2 /A 176 4变异 2 6例 ,两组相比 ,差异无显著性 (P >0 0 5 )。同时对 15例中、重型慢性乙型肝炎病人血清检测 ,发现T 176 2 /A 176 4变异11例 ,在 5例发生变异株与野生株的混合感染中 ,变异株有优势积累趋势。结论　HBVC基因启动子变异特别是T 176 2 /A 176 4联合位点变异在我国慢性乙型肝炎患者中普遍存在 ,变异的发生只是降低了HBV前CmRNA的转录效率 ,而不是完全中止HBeAg的复制 ,这种变异明显不同于前C区终 2 8变异对血清学的影响 ,且随着病情的发展 ,变异株发生优势积累。提示 ,这一变异似乎与病理变化轻重程度有一定关系。Objective To study the core promoter mutation in chronic hepatitis B and its effect on viral serology.Methods HBV core promoter gene fragments were amplified by using mismatched PCR combined with a restriction fragment length polymorphism assay. The PCR products were digested with BcL I and subjected to electrophoresis on agarose gels.Results We investigated the core promoter mutation in 89 patientswith HBV DNA positive. The patterns of restriction fragment length polymorphism (RFLP) of the core promoter gene were distinguished and verified by direct sequencing. The combined mutations of nucleotides (nt) 1762 and 1764 in the core promoter from A to T and G to A were detected in 47 individuats, 21 of 43 HBeAg positive patients and 26 of 46 anti HBe positive cases were found infected with this combined mutant. These two groups showed no significant difference (P>0.05).The combined mutation was found in 11 of the 15 patients with medium and severe chronic hepatitis. In 5 chronic hepatitis B patients with coexistence of wild type and mutant a tendency of superior accumulation of mutant was found.Conclusion This study suggests that the core promoter mutations commonly exist in chinese chronic hepatitis B patients. The mutation can only reduce the pre core mRNA transcribing efficiency, but can not discontinue the synthesis of HBeAg. The effect on serology in this mutation is different from that in pre core stop 28 mutation. The superior accumulation of mutations seems relating to the degree of chronic liver disease.

关 键 词：乙型肝炎病毒 变异 RFLP C基因 启动子 检测 

分 类 号：R446.5[医药卫生—诊断学]