胆囊结石病人线粒体高变区测序结果分析  

Sequence analysis of hypervariants of mitochondrial DNA in patients with cholelithiasis

作　　者：张宇[1] 牛振民 韩天权[1] 秦俭[1] 姜志宏[1] 张凯悦 黄薇 张圣道[1] 

机构地区：[1]上海第二医科大学附属瑞金医院外科上海消化外科研究所,上海市200025 [2]中国人类基因组南方研究中心

出　　处：《中华肝胆外科杂志》2005年第9期580-582,共3页Chinese Journal of Hepatobiliary Surgery

基　　金：国家自然基金资助项目(基金编号30271272)

摘　　要：目的检测胆囊结石病人和正常对照个体线粒体DNA第一高变区(HVS1)单核苷酸多态性(SNPs),分析胆囊结石病的可能易感因素。方法对上海地区96个来自不同家系的胆囊结石病人和204例正常对照个体进行线粒体DNA第一高变区序列检测,将测序结果与标准序列对比,寻找存在的单核苷酸多态性,并据此进行线粒体单倍型类群划分。比较单倍群在两组之间的分布差异,估算各种单倍群人群胆囊结石病相对发病风险。结果B单倍群、B4和B4b1亚群在胆囊结石病组的出现频率明显高于对照组(P<0·05);胆囊结石病组和正常对照组B单倍群的比值比(OR)为2·005;B4亚群OR值为2·200;B4b1亚群OR值为3·9331,P均<0·05。结论线粒体DNA B4b1单倍型亚群是胆囊结石病的易感因素。Objective To sequence the mitochondrial DNA hypervariants in patients with cholelithiasis and healthy individuals to find their single nucleotide polymorphisms （SNPs） and investigate the predisposing factors resulting in cholelithiasis. Methods Genomic DNA was extracted from the whole-blood samples from 96 patients of different cholelithiasis pedigrees and 204 irrelative controls living in Shanghai, China. Mitochondrial DNA （mtDNA） hypervariant 1 （HVS1） was chosen as the target fragment and amplified by PCR. The electrophoresis of PCR products was performed to test their purity and then the products were directly sequenced with the same primer as that used for PCR amplification on 3730 Sequence Detection System. The sequence was edited and aligned by use of the Chromas 2.23 software and BLAST and the mutation was scored based on the revised reference sequence （rCRS） （Andrews 1999）. According to the HVS1 motif of every individual, the mtDNAs were classified into haplogroups. The frequency of each haplogroup was compared by Chi-square test between the patients with cholelithiasis and controls to investigate the distribution difference. Meanwhile, the odds ratio （OR） was determined. Results There were significant differences in frequencies and ORs of haplogroups B, B4 and B4bl between the patients with cholelithiasis and controls （P〈 0. 05）. The OR of B4bl was the highest and its 95% confidence interval was 2.17-12.71. Conclusions In south China, people of the B4bl haplogroup are predisposed to cholelithiasis.

关 键 词：胆结石 线粒体DNA 易感性 单倍群 胆囊结石病 第一高变区 病人 测序 单核苷酸多态性 正常对照组 

分 类 号：R575.6[医药卫生—消化系统]