快速简便筛检乙型肝炎病毒C基因启动子变异的方法及应用

目的明确中国乙型肝炎病毒（ＨＢＶ）Ｃ基因启动子（ＢＣＰ）区段变异特点及其可能的临床意义。方法应用错配聚合酶链反应限制性片段长度多态性（ＰＣＲ－ＲＦＬＰ）方法，结合核苷酸序列分析，检测１４３例ＨＢｓＡｇ阳性的ＨＢＶ慢性感染者中Ｘ基因／ＢＣＰ区的核苷酸（ｎｔ）１７６２碱基由Ａ→Ｔ和１７６４碱基由Ｇ→Ａ变异。结果在１１４份ＨＢＶＤＮＡ阳性血清中，３７例感染ＢＣＰ变异株，其中３１例为慢性肝炎病例，６例为慢性ＨＢＶ无症状感染者（ＡｓＩ）；其中４１例ＨＢｅＡｇ阳性ＡｓＩ中仅２例检出，１８例ＨＢｅＡｇ阴性ＡｓＩ中也有４例检出。ＨＢｅＡｇ阳性的６５例中，１３例有ＢＣＰ区变异，而ＨＢｅＡｇ阴性的４９例中，２４例有ＢＣＰ区变异（Ｐ＜０．０１）。结论错配ＰＣＲ－ＲＦＬＰ检测这一对点突变，具有快速、简便、适用的优点；ＨＢＶ毒株ＢＣＰ变异可能与肝病变活动有一定关系。

Detection of core promoter sequence mutations of hepatitis B virus by a rapid and simple method

Objective In order to explore the clinical significance and epidemiological characteristics of the variant, we investigated the prevalence of the core promoter variant in 114 Chinese HBV isolates. Methods Mismatched PCR combined with a restriction tragment length polymorphism assay was used in the study. Results The combined mutation of nucleotides(nt)1762 and 1764 in the core promoter from A to T and G to A was detected in 37 individuals, 31 of which suffered with advanced liver diseases and the other 6 were asymptomatic carriers. On the other hand, 13 of 65 HBeAg positive patients or asymptomatic carriers and 24 of 49 HBeAg negative cases were found infected with the mutant, respectively. The incidence of core promoter mutant was lower in HBeAg positive asymptomatic carriers or patients suffered with hepatitis B than those HBeAg negative ones. Conclusion This method for rapid and simple detection of the combined mutation in core promoter should be useful for evaluation of change of core promoter variant population during the course of HBV infection. This study suggest that the core promoter mutations might result in decreased HBeAg synthesis and lead to persistent infection of HBV.