乙肝肝硬化患者HBV前C区和基本核心基因启动子序列分析

Sequence analysis of precore and core promoter of HBV isolated from patients with liver cirrhosis

用套式PCR(nPCR)对 3 5例肝硬化患者血清HBV前C区和基本核心基因启动子 (BCP)进行扩增 ,阳性者用直接测序法进行序列分析。结果 2 3例HBVDNA阳性 ,阳性率为 65 7% ( 2 3 / 3 5 ) ,无正常序列标本 ,与e抗原产生有关的突变 :nt1762、1764双突变 (AT、GA) ,占 73 9% ( 17/ 2 3 ) ;nt1896点突变 (GA) ,导致终止密码产生 ,占2 1 7% ( 5 / 2 3 ) ;nt185 8点突变 (TC) ,占 2 1 7% ,其中标本 417、42 6、43 0同时在nt 185 6发生点突变 (CT)。其它的突变有nt1799点突变 (CG) ,占 47 8% ( 11/ 2 3 ) ,为无义突变 ;有 6例在nt1846发生点突变AT ,4例nt180 2 - 180 4发生突变 (TTCCGT) ;4例nt175 2位点突变 (AG) ;标本 43 2在nt175 1插入TG导致移码突变 ,并在nt1774- 1874发生缺失突变。说明HBVBCP和前C突变株在肝硬化患者中很常见 。

The nested polymerase chain reaction (nPCR) was used for amplification of HBV DNA core promoter and PreC in 35 sera form patients with liver cirrhosis in Guangxi,and then HBV DNA nPCR products were sequenced by direct sequencing.The results show that HBV DNA positive rate of 35 patients is 65.7%(23/35).All patients were infected with mutant.The double mutation (AT at nt1762 and GA at nt 1764)affecting the producing of HBeAg counts for 73.9%(17/23).And the mutation resulting in stop code at nt 1896 counts for 21.7%(5/23).Mutation at nt1858(TC)counts for 21.7%,of which mutation at nt 1856(CT) was seen in samples 417,426 and 430.The other mutation such as AG at nt 1752,CG at nt1799,AT at nt1846,TTCCGT at nt 1802-1804 count for 17.4%,47.8%,26.1% and 17.4% respectively.The mutation that TG insert between nt 1751-1752,then delete between 1774-1874 was seen in sample 432.The results suggested that HBV core promoter and preC mutations prevalent among patients with liver cirrhosis.These mutations are associated with the development of liver cirrhosis.