关键词: 肝豆状核变性, ATP7B基因, 聚合酶链反应, 高分辨熔解曲线分析, 产前基因诊断

Abstract: Objective To identify the ATP7B mutation and perform the prenatal gene diagnosis in a family with Wilson’s disease(WD). Methods Genomic DNA was extracted from the peripheral blood and the fetal villi tissue by standard phenol/chloroform method. PCR and Sanger DNA sequencing was used to analyze the causative mutation in the proband; Mutation identification was performed in the other 4 familial members including a fetus with high risk of WD by PCR-HRM. In the same way, a prenatal gene diagnosis was completed successfully in the mother of the proband. DNA sequencing was used to validate the genotype of the samples in different HRM-curves. Results Genetic sequencing showed that the proband was a homozygote of the missense mutation c.2333G>T (p.R778L) , which was located in the exon 8 of gene ATP7B. HRM analysis displayed three different curves, representing three different genotypes. The curve types for the 5 familial members and 4 normal control samples were consistent with genetic sequencing: the proband himself was a mutant homozygote, all the other family members were the heterozygote of the mutation, and the 4 normal controls were wild type. Conclusions The authors found a known mutation of p.R778L in a WD family, and the prenatal gene diagnosis was achieved in a fetus with high risk by PCR-HRM.

Key words: Wilson’s disease, ATP7B gene, polymerase chain reaction, high melting resolution, prenatal gene diagnosis