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目的 动态研究中国人群HCVⅡ /1b型包膜蛋白E2 /NS1高变区 1(HVR1)序列变异规律、意义及影响因素。方法 应用逆转录巢式PCR技术从 2 0例HCVⅡ /1b型感染的中国病人血清中扩增了HCV部分包膜区基因片段 (nt144 9～ 15 86 ,HCV J) ,纯化后直接采用双脱氧链末端终止法进行序列分析。结果 中国人群HCVHVR1位于氨基酸 (AA) 384～ 410位 ,有 6个较保守的AA位点 :385位Thr,389、390、40 6位Gly ,40 3位Phe ,40 9位Gln。自然病程组核苷酸 (nt)变异率每年 [0～ 2 0 7(平均0 5 0 ) ]× 10 1个 /位点 ,AA变异率每年 [0～ 6 2 2 (平均 1 2 4) ]× 10 1个 /位点 ,干扰素治疗组nt变异率为每年 [0～ 5 0 3(平均 2 31) ]× 10 1个 /位点 ,AA变异率每年 [0～ 10 7(平均 4 40 ) ]× 10 1个 /位点。干扰素治疗对HVR1区变异有较强的免疫选择作用 ,HVR1变异率高低与慢性肝炎的临床病程有一定联系 ,肝炎活动期变异明显。结论 对HCVHVR1区变异规律及其生物学意义的进一步研究将有助于了解HCV慢性感染机理、制定新的治疗方案及疫苗设计等。
Objective To study the variation of HVR1 sequence and its influencing factors in HCV Ⅱ / 1b envelope protein E2 / NS1 in Chinese population. Methods Reverse transcriptase nested polymerase chain reaction (PCR) was used to amplify HCV partial envelope region (nt144 9 ~ 1586, HCV J) from 20 Chinese patients with HCV Ⅱ / 1b infection. The purified DNA was directly purified by dideoxy chain Terminal termination method for sequence analysis. Results HCVHVR1 in Chinese population was located at amino acid (AA) 384-410, with six conserved AA sites: 385 Thr, 389,390, 40 6 Gly, 40 3 Phe and 40 9 Gln. The variation rate of nucleotide (nt) of natural disease group was from 0 to 270 (average 0 0) per 100 × 10 1 loci per year. The variation rate of AA was from 0 to 6 2 2 (average 12 4) per year. × 10 1 / locus. The variation rate of nt in interferon treatment group was [0 ~ 500 (average 2 31)] × 10 1 loci per year. The variation rate of AA was between 0 ~ 10 7 per year )] × 10 1 per locus. Interferon treatment has a strong immunosuppressive effect on HVR1 mutation. The mutation rate of HVR1 is related to the clinical course of chronic hepatitis, and the variation of hepatitis activity is obvious. Conclusion The further study of the variation of HCVHVR1 and its biological significance will help to understand the mechanism of chronic infection of HCV, to develop new treatment options and vaccine design.