江苏徐州地区肝豆状核变性患者ATP7B基因8、13号外显子突变的检测和分析

目的研究徐州地区肝豆状核变性(Hepatolenticular degeneration,HLD)患者ATP7B基因8、13外显子突变情况,为本病的早期和产前诊断提供理论依据。方法采集33例HLD患者和30例对照组正常人外周血、提取DNA、PCR扩增ATP7B基因8和13外显子;对扩增产物分别进行限制性内切酶MspI及BtgI酶切分析,反应异常者行DNA测序,最后将突变结果与临床表型作相关性分析。结果 33例HLD患者中,15例存在MspI酶切异常,测序为Arg778Leu杂合或纯合突变,占45.45%(15/33);9例存在BtgI酶切异常,测序为Pro992Leu杂合突变,占27.27%(9/33)。30例正常对照组未检测出突变。结论 ATP7B基因第8、13外显子是徐州地区HLD患者的基因突变热区,筛选本地区HLD可疑患者时应优先检测。

Detection and analysis of the mutation in exon 8 and 13 of ATP7B gene in patients with hepatolenticular degeneration located in Xuzhou

Objective:To investigate the mutation of exon 8 and 13 of ATP7B gene in patients with hepatolenticular degeneration(HLD) located Xuzhou of China,based prenatal diagnosis of early diagnosis and provide a theoretical basis.Methods: Extract the genomic DNA from 33 HLD patients and 30 normal controls,and amplify exon 8,13 of ATP7B gene by polymerase chain reaction(PCR).The amplification products of exon 8 and exon 13 were digested with MspI and BtgI respectively followed by sequencing the PCR products of exon 8 and 13 from all the patients and normal controls.Results: Digested by MspI through amelioration,15 cases were abnormal.Sequence results showed that 45.45%(15/33)of the cases had homozygous or heterozygous Arg778Leu mutation in exon 8.9 cases were abnormal using digestion by BtgI,direct sequencing showed 27.27%(9/33)of the cases had heterozygous Pro992Leu mutation in exon 13.No abnormality was found in normal controls group.Conclusion: Arg778Leu in exon 8 and Pro992Leu in exon 13 are the hot points mutation in HLD patients which come from Xuzhou of China,and they are the exons which should be detected preferentially when screening doubtful HLD patients.