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目的检测湖南地区汉族人群肝豆状核变性患者(WD)ATP7B基因常见突变种类和形式。方法提取22例WD患者外周血基因组DNA,聚合酶链反应(PCR)扩增ATP7B基因第5、8、12及13号外显子并进行DNA直接测序检测,应用在线BLAST软件分析。结果22例患者中共发现15例患者存在基因突变。其中10例患者存在8号外显子2273G→T杂合突变(即Arg778Leu),且均伴有2250C→G多态(即Leu770Leu,均为杂合子),未发现纯合突变。12号外显子中共发现2855G→A多态(即Arg952Lys)7例(杂合突变4例,纯合突变3例),其中1例合并12号外显子2828G→A杂合突变(即Gly943Asp),另3例合并Arg778Leu杂合突变。13号外显子2975C→T杂合突变(即Pro992Leu)1例。5号外显子未发现突变。结论Arg778Leu是湖南地区汉族WD患者的突变热点,5号外显子为非突变热点。
Objective To detect the common mutation types and forms of ATP7B gene in patients with hepatolenticular degeneration (WD) in Han nationality in Hunan Province. Methods Genomic DNA was extracted from peripheral blood of 22 cases of WD patients. Exon 5, 8, 12 and 13 of ATP7B gene were amplified by polymerase chain reaction (PCR) and sequenced by DNA direct sequencing. The online BLAST software was used to analyze the genomic DNA. Results A total of 15 patients in 22 patients were found to have gene mutation. Among them, there was a 2273G → T heterozygous mutation (ie, Arg778Leu) on exon 8 in 10 patients. All of them were associated with 2250C → G polymorphism (ie, Leu770Leu, all heterozygous). No homozygous mutation was found. There were 7 cases of 2855G → A polymorphism (ie, Arg952Lys) in exon 12 (4 cases of heterozygous mutation and 3 cases of homozygous mutation), of which 1 case had 2828G → A heterozygous mutation (Gly943Asp) on exon 12, The other 3 cases combined Arg778Leu heterozygous mutation. Exon 13 2975C → T heterozygous mutation (ie Pro992Leu) in 1 case. No mutation was found in exon 5. Conclusion Arg778Leu is a hot mutation in WD patients in Han nationality in Hunan Province. Exon 5 is a non-mutated hot spot.