Non-sequencing molecular approaches to identify preS2-defective hepatitis B virus variants proved to be associated with severe liver diseases
Non-sequencing molecular approaches to identify preS2-defective hepatitis B virus variants proved to be associated with severe liver diseases
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DOI：
10.1016/j.jhep.2003.11.025
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发表时间：
2004-03-01
影响因子：
25.7
通讯作者：
Brancatelli, S
中科院分区：
文献类型：
--
作者：
Raimondo, G;Costantino, L;Brancatelli, S
背景/目的：慢性乙型肝炎病毒（HBV）感染过程中可能出现PreS 2缺陷型B病毒变异。这些变体在ATG起始密码子处携带突变和/或在preS 2基因组区域中携带框内缺失，并且通常通过测序分析来检测。我们评估了这些变异的患病率在一个大系列的慢性HBV感染patients通过非测序molecular approaches.Methods：我们检查了HBV分离株从110 HBV携带者：15个是非活动载体（IC），50慢性肝炎（CH），25是approaches，19有肝细胞癌（HCC）。PCR扩增前S2基因组全序列。处理扩增子：（A）通过丙烯酰胺凝胶电泳显示缺失的基因组;（B）通过经Nla III酶消化后的琼脂糖凝胶电泳，所述Nla III酶切割野生型ATG起始密码子而不切割突变型密码子。特别地，我们在2/15 IC、25/50 CH、13/26 APC和16/19 HCC中发现了preS 2缺陷突变体。因此，这些变异的存在与活动性感染和肝脏疾病显著相关（P < 0.002）。结论：我们的非测序分子生物学方法灵敏度高、特异性强，可简化HBV前S2变异型的鉴定。这些变异体的感染与活动性感染和HCC显著相关。(C)2003年欧洲肝脏研究协会。Elsevier B. V.出版，保留所有权利。
Background/Aims: PreS2-defective hepatitis B virus (HBV) variants may emerge during chronic HBV infection. These variants carry mutation(s) at the ATG-start-codon and/or in-frame deletion into the preS2 genomic region and are commonly detected by sequencing analyses. We evaluated the prevalence of these variants in a large series of chronic HBV infected patients through non-sequencing molecular approaches.Methods: We examined HBV isolates from 110 HBV carriers: 15 were inactive carriers (IC); 50 had chronic hepatitis (CH); 25 were cirrhotics; 19 had hepatocellular carcinoma (HCC). The entire preS2 genomic region was amplified by PCR technique. The amplicons were processed: (A) through electrophoresis on acrylamide gel to reveal deleted genomes; (B) through electrophoresis on agarose gel after digestion by Nla III enzyme that cuts the wild ATG-start-codon but not the mutated one.Results: We detected preS2 variants in 56/110 cases (51%). In particular, we found preS2-defective mutants in 2/15 IC, 25/50 CH, 13/26 cirrhotics, and 16/19 HCC. The presence of these variants was thus significantly associated with active infection and liver disease (P < 0.002). Moreover, among cases with liver disease preS2-mutants were more prevalent in HCC patients (P < 0.02).Conclusions: Our non-sequencing molecular methods are sensitive and specific, and simplify the identification of all preS2 HBV variant forms. Infection by these variants is significantly associated with active infection and HCC. (C) 2003 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.