与ATP7B基因紧密连锁的微卫星DNA单体型在汉族Wilson病的多态性分析

Polymorphism of ATP7B Related Microsatellite DNA Haplotypes in the Ethnic Han Chinese with Wilson Disease

摘要:

目的：了解与ATP7B基因紧密连锁的微卫星DNA(D13S314，D13S301和D13S316)单体型在正常人、Wilson病(WD)患者及其杂合子的分布特点及意义。方法：采用荧光标记3个短串联重复标记(D13S314，D13S301和D13S316)，测序定位和Genotype TM 软件技术分析71例WD患者和123位父母的单体型。结果：在D13S314，D13S301和D13S316位点分析中得到71例WD患者和123位携带者及54例正常个体等位基因片段；片段大小分别为134～157 bp，128～156 bp 和136～154 bp；获得等位基因数分别为19，20和15个；3个位点的杂合率分别是 0.79，0.82 和 0.23。D13S314，D13S301和D13S316位点的等位基因分布在WD患者和正常人群之间明显不同，其中D13S314和D13S301位点显示各有9个等位基因片段存在明显差异(P<0.05)，D13S316位点显示有4个等位基因片段存在明显差异(P<0.01)；显示的81种单体型中以 12-6-5，15-10-5，6-10-5和6-14-5最多见，分别占 5.2%，4.5%，4.5% 和 3.7%，其次为12-8-5，12-9-5和6-16-5，各占 3.0%，13-10-8，6-13-5，6-14-13和6-9-5各占 2.2%。结论：单体型的类型较多可能和基因突变的类型多样化相关，D13S314-D13S301-D13S316的单体型分析对WD的诊断，尤其是症状前或产前诊断都有重要的意义，是有效及可行的方法之一。

Abstract:

OBJECTIVE: To explore the distribution and significance of haplotypes of microsatellite DNA (D13S314, D13S301 and D13S316 ) closely related to ATP7B in normal population, Wilson disease (WD) patients and heterozygotes. METHODS: Using three well characterized short tandem repeat markers of fluorescence labelling (D13S301, D13S314 and D13S316), localization and sequence were studied with Genotype TM software in 71 WD patients and 123 WD parents. RESULTS: Based on the analysis of haplotypes of D13S314, D13S01 and D13S316, 19, 20 and 15 alleles were obtained in the 71 patients with WD, 123 carriers and 54 normal persons, respectively. The size of segments was 134 157 bp, 128 156 bp, and 136 154 bp, respectively. Heterozygosity was 0.79 , 0.82 and 0.23 , respectively. There was significant difference in the distribution of alleles of D13S314, D13S301 and D13S316 makers between the WD patients and normal persons (9 alleles were noted in D13S314 maker, 9 alleles in D13S301 maker and 4 alleles in D13S316 maker)(< 0.05 or 0.01 ). There were 81 haplotypes. Of them, 12 6 5, 15 10 5, 6 10 5 and 6 14 5, the commonest haplotypes, accounted for 5.2% , 4.5% , 4.5% and 3.7% respectively; 12 8 5, 12 9 5 and 6 16 5 separately accounted for 3.0% ; 13 10 8, 6 13 5, 6 14 13 and 6 9 5 separately accounted for 2.2% . CONCLUSIONS: The complexity of haplotypes may be related to the complexity of mutation spectrum of ATP7B. The haplotype analysis of D13S314 D13S301 D13S316 is very valuable in making a diagnosis of WD, especially in presymptomatic patients. It is also useful for prenatal diagnosis of WD.