13例Wilson病患者基因8号外显子778密码子碱基突变的SSCP及酶切分析

Gene Analysis of 13 Cases of Wilson Disease With SSCP and Enzymolysis: Alkal Mutation of 778 Code in 8th Exon

目的 寻找山东籍Wilson病患者WND基因突变热点,以便易于用SSCP及酶切分析方法检出。使无先证者及独生子女家系易于作出基因诊断,同时可用于杂合子筛查。方法 PCR扩增WND基因8号外显子778密码子区域,SSCP检洲点突变,用MSPI内切酶进一步验证G-T突变的正确性。结果 13例Wilson患者,检出778密码子G-T纯合突变2例,杂舍突变7例,未突变4例,共计26条染色体,突变染色体11条,总染色体突变率为42.3％。结论 WND基因8号外显子,778密码子G-T突变是山东WD患者的一个突变热点。SSCP和酶切分析是诊断此位点突变的便捷方法,可用于WD患者及杂合子检出。

Objective To examine WND gene mutation point in WD patients in shandong area. So as to use SSCP and enzymolysis to make gene diagnosis. Especially in no proband family or one child family. It's also used to screen hybrid. Method 778 code region in 8th exon in WND gene was amplyfied with PCR. Point mutation was identified with SSCP. The G-T mutation was proved with MSPI endonuclease analysis. Result In 13 cases WD there're 2 778 code G-T purebrid mutation, 7 hybrid and 4 none mutation. There're 26 chromosome in total. Of which 11 were mutation. All chromosome mutation was 42. 3 per cent. Conclusion 778 code G-T mutation of 8th exon in WND gene is a mutation point in WD patients in Shandong area. SSCP and enzymolysis is a easy and quick method to diagnose this mutation. It can also be used to screen WD hybrids.