234例慢性乙型肝炎患者HBV前C/BCP区突变分析 |
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引用本文: | 白丽,王琳,李乐,孟玉华,万芃瑾,刘晓红,陈雪源,张玲霞,徐东平. 234例慢性乙型肝炎患者HBV前C/BCP区突变分析[J]. 实用肝脏病杂志, 2009, 12(1): 14-16,40 |
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作者姓名: | 白丽 王琳 李乐 孟玉华 万芃瑾 刘晓红 陈雪源 张玲霞 徐东平 |
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作者单位: | 解放军第三○二医院传染病研究所病毒性肝炎研究室,北京市,100039 |
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基金项目: | 国家重点基础研究发展规划(973计划) |
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摘 要: | 目的调查慢性乙型肝炎患者血清HBV前C/基本核心启动子(BCP)区的突变情况,分析各种突变对HBeAg表达的影响。方法抽提患者血清DNA,采用改进的巢式PCR技术,扩增HBV前C/BCP区基因,对PCR产物进行DNA测序。用NTI软件比对结果,根据文献报道,选取1753、1762、1764、1862、1896和1899共6个位点进行突变分析,并重点分析不同突变对患者血清HBeAg阳性率和病情的影响。结果在234例中6个位点均无突变者为74例(31.6%),其血清HBeAg阳性率为74.3%(55/74)。在234例中6个位点检出至少1种突变的病例为160例(68.4%),突变形式包括4种单一位点突变和21种组合形式的突变;检出G1896A突变73例(31.2%),其中36例检出有共存未突变序列,37例仅检出突变序列,后者血清HBeAg的阳性率为18.9%(7/37)。检出G1896A以外其他形式突变的有87例,HBeAg阳性率为63.2%(55/87);其中以A1762T+G1764A最为常见,HBeAg阳性率为69.4%(34/49)。在1753、1862位点上检出4种特殊碱基突变,前C区有基因片段插入或缺失的有2例。结论多数慢性乙型肝炎患者在HBV前C/BCP区可检出突变,突变形式多样,其中G1896A突变样本血清HBeAg阳性率显著下降,而其他突变对其影响较小。应用DNA序列测定法分析HBV前C/BCP区基因突变,获得的信息全面,对临床评估病情进展和实施抗病毒治疗有参考价值。
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关 键 词: | 乙型肝炎 HBV前C/BCP区突变 HBeAg 基因测序 |
Mutation of hepatitis B virus precore/basal core promoter in 234 patients with chronic hepatitis |
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Affiliation: | BAI Li, WANG Lin,Ll Le,et al. (Viral Hepatitis Research Laboratory,Institute of Infectious Diseases,302 Hospital of PLA ,Bering 100039, China) |
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Abstract: | Objective To investigate the occurrence of hepatitis B virus (HBV)precore/basal core promoter(BCP) mutations and its correlation on serum HBeAg expression in patients with chronic hepatitis B(CHB). Methods Sera were collected for extraction of HBV DNA. The precore/BCP gene fragment was amplified by an in-house nested PCR assay and analyzed by direct PCR-produet sequencing. Based on reference literatures,6 mutation sites which possibly influencing HBeAg expression,secretion,or HBV replication were selected as follows:1753,1762,1764,1862,1896 and 1899. The data were processed by NTI software. Serum HBeAg was detected by enzyme-linked immunosorbent assay. Results Seventyfour (31.6%) of 234 patients enrolled in the study were mutation-negative at all 6 sites. Serum HBeAg positive rate of these cases was 74.3%. One hundred and sixty (68.4%) of the 234 patients were detected with various mutations at the sites,including 4 single-site and twenty-one multiple-site mutations. Among the total,73(31.2%) cases contained G1896A with or without coexistence of wild-type sequence at the sites,of whom 37 cases without coexistent wild-type sequence at the site had 18.9% HBeAg positive rate. Preeore/BCP mutations other than G1896A were detected in 87 patients whose HBeAg positive rate was 63.2%(55/87). A1762T+G1764A was the most frequent mutation in patients positive with other than G1896A mutations,and the positive HBeAg was documented in 69.4%(34/49) of these patients. Four unusual substitutions were detected at site 1753 and 1862. Two cases were detected with insertion or deletion in precore/BCP region. Conclusion HBV precore/BCP mutations exist in most CHB patients in diverse forms. The patients with G1896A mutation had a significantly lower positive rate of serum HBeAg,while other mutations detected had little influence on HBeAg secretion. Screening HBV precore/BCP mutations by DNA sequencing could obtain rich information and thus is valuable for evaluating disease progression and administrating anti-viral therapy in clinical practice. |
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Keywords: | HBeAg |
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