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目的研究慢性乙型肝炎病毒(HBV)感染者中HBV基因型C亚型(HBV/C)的核心启动子、前C/核心区基因变异情况,分析HBV/C亚型的病毒学特征。方法用酶联免疫法(ELISA)筛选出79例HBV/C,再用聚合酶链反应-限制性酶长度多态性分析方法(PCR-RFLP)进行HBV/C亚型分析;同时针对HBV核心启动子、preC/核心区基因进行半巢式PCR及PCR产物直接测序。结果①79例HBV/C中,33例(41·8%)为HBV/C1亚型,46例(58·2%)为HBV/C2亚型。②HBV/C1亚型仅见于来自中国南方的患者(P<0·0001)。③A1898位点变异仅见于HBV/C1亚型(P=0·056),V1753位点变异在HBV/C1亚型中多见(P<0·05);HBV/C2以T1858(90%)、A1896(40%)位点变异多见(P<0·008)。T1762/A1764位点变异在HBV/C两种亚型中均常见。④肝细胞癌(HCC)患者中,V1753和T1762/A1764变异最常见(P<0·05)。结论HBV/C1和HBV/C2在中国有明显的地区差异;V1753合并T1762/A1764双变异与发展为HCC相关,尤其在HBV/C1患者。
Objective To study the variations of the core promoter and precore / core gene of HBV genotype C subtype (HBV / C) in patients with chronic hepatitis B virus infection and to analyze the virological characteristics of HBV / C subtypes. Methods 79 cases of HBV / C were screened by enzyme-linked immunosorbent assay (ELISA) and HBV / C subtypes were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) Promoter, preC / core region genes were subjected to semi-nested PCR and PCR products were sequenced directly. Results ① Of the 79 cases of HBV / C, 33 (41.8%) were HBV / C1 subtypes and 46 (58.2%) were HBV / C2 subtypes. ② HBV / C1 subtype only found in patients from southern China (P <0.0001). (3) The mutation of site A1898 was found only in HBV / C1 subtype (P = 0.056). The variation of site V1753 was more common in HBV / C1 subtype (P <0.05) A1896 (40%) locus variation more common (P <0 · 008). T1762 / A1764 site variation in HBV / C are common in both subtypes. Among the patients with hepatocellular carcinoma (HCC), the variations of V1753 and T1762 / A1764 were the most common (P <0.05). Conclusion HBV / C1 and HBV / C2 have obvious regional differences in China. The double mutation of V1753 combined with T1762 / A1764 is associated with the development of HCC, especially in HBV / C1 patients.