江苏徐州地区肝豆状核变性患者ATP7B基因12、14、18号外显子的检测和分析

Detection and analysis of the mutation of exon 12,14,18 of Wilson's disease gene iocated in Xuzhou

目的研究江苏徐州地区肝豆状核变性(又称Wilson'病,WD)患者ATP基因12、14、18外显子突变情况,以期建立徐州地区WD的ATP基因突变热点的检测平台。方法采集45例WD患者及50例正常人外周血,提取基因组DNA,PCR扩增ATP7B第12、14、18号外显子,12号外显子扩增产物进行Tail内切酶酶切分析;然后对所有病人及正常人DNA外显子扩增产物进行直接测序,进而与临床表型做相关分析。结果 45例WD病人中,共发现6例突变。12号显子无1例存在Tail酶切反应异常,测序共发现2855G→A多态(即Arg952Lys)9例,其中1例合并12号外显子2828G→A杂合突变(即Gly943Asp),18号外显子发现3884C→T(即Ala1295Val)杂合突变5例,其中2例合并3889G→A多态(即Val1297lle),14号外显子未见突变。结论 18号外显子可能是徐州地区WD患者的基因突变热区,可作为徐州地区WD可疑患者的优先筛选区域之一。12、14号外显子可能不是徐州地区WD患者的基因突变热区。

Obiective： To sequence and study the exon 12,14,18 of hepatolenticular degeneration gene（Wilson＇s disease,WD） located Xuzhou of China,and to establish the ATP gene mutation hot test platform.Methods：Extract the genomic DNA from 45 WD patients and 50 normal controls,and amplify exon 12,14,18 of ATP7B gene by polymerase chain reaction（PCR）.The amplifi cation products of exon12 were digestcd with Tail respectively followed by sequencing the PCR products of exon 12,14,18 from all the patients and normal controls.The correlation between the mutation and clinical manifestation were stucdied.Results：six of the 45 patients were found with mutations.Digested by Tail through amelioration No case was abnormal.Direct sequence result shows that nine patients were found with 2855G→A（Ar9952Lys） polymorphism,one of them had 2828G→A（Gly943Asp） heterozygous mutation in exon 12,five patients were found with 3884C-T（Ala1295Val）heterozygous mutation,two of them were found with 3889G-A（Val1297lle） polymorphism in exon 18,No mutations were found in exon 14.Conclusions： Exon 18 of ATP7B gene maybe is the hot points of WD genetic mutation in the population of Xuzhou,and maybe it is the exons which should be detected preferentially when screening doubtful WD pationts.exon 12 and exon 14 of ATP7B gene maybe is not the hot points of WD genetic mutation in the population of Xuzhou.