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目的:通过对经母婴传播获得HBV感染的子女及其母亲慢性携带者体内HBV EnhII/CP/PreC基因序列研究,了解来源相同的HBV毒株在不同程度病毒血症情况下EnhII/CP/PreC基因变异特点。方法:选择15对母子AsC,根据HBV病毒血症高低分为3组,每组5对母子,即Ⅰ组(母子均为高病毒血症)、Ⅱ组(子女为高病毒血症、母亲为低病毒血症)和Ⅲ组(子女为低病毒血症、母亲为高病毒血症)。同对母子HBV亚型相同,各组中有4/5对母子为B/adw2、1/5对母子为C/adrq+亚型。应用T-A克隆技术构建重组质粒pGEM-EnhII/CP/PreC、双酶切鉴定,每个病人选2个酶切鉴定正确的克隆测序并分析。结果:高病毒血症组间或低病毒血症组间HBV EnhII/CP/PreC基因变异数目及位点均无明显差异,低病毒血症病人变异数目及位点明显高于高病毒血症。高病毒血症中16例B/adw2亚型的32个EnhII/CP共有6个变异位点;4例C/adrq+亚型的8个克隆中有4个变异位点,均无热点变异;PreC无变异。低病毒血症中8例B/adw2亚型病人的16个克隆共有26个变异位点,变异与年龄无关。结论经母婴传播获得HBV感染儿童及其母亲无症状携带者中,HBVEnhII/CP/PreC变异与病毒血症高低有关,低病毒血症病人变异明显高于高病毒血症。HBV的低复制状态是CP、EnhII及X蛋白等结构和功能改变综合影响的结果,与年龄无关。
OBJECTIVE: To study the sequence of HBV EnhII / CP / PreC gene in HBV carriers and their mothers who had been infected by mother-to-child transmission, and to understand the effect of EnhII / CP / PreC Gene mutation characteristics. Methods: Fifteen pairs of mother and child AsC were selected and divided into 3 groups according to the level of HBV viremia. Each group consisted of 5 pairs of mother and child, group Ⅰ (mother and child were all viremia), group Ⅱ (children with viremia, Low viremia) and Group III (low viremia in children and high viremia in the mother). The same as the mother and child HBV subtypes, each group 4/5 pairs of mother and child B / adw2, 1/5 pairs of mother and child C / adrq + subtype. The recombinant plasmid pGEM-EnhII / CP / PreC was constructed by T-A cloning technique. The double digestion and restriction enzyme digestion were performed. Two clones were selected for each patient to identify the correct clones and analyze them. Results: There was no significant difference in the number of HBV EnhII / CP / PreC genes between viremia group and low viremia group. The number of viremia and locus in patients with low viremia was significantly higher than that of viremia. There were 6 mutation sites in 32 EnhII / CPs of 16 B / adw2 subtypes in viremia; 4 of 8 clone sites in 4 C / adrq + subtypes had no hot spot mutation; and PreC No variation. There were 26 variation loci in 16 clones of 8 cases of B / adw2 subtype in low viremia, and the variation was not related to age. Conclusion The variation of HBVEnhII / CP / PreC in HBV-infected children and their mothers with asymptomatic carriers is related to the level of viremia. The variation of patients with low viremia is obviously higher than that of viremia. The low replication status of HBV is the result of a combination of changes in the structure and function of CP, EnhII, and X proteins, regardless of age.