Analysis of The Mutation of Exon 8、12、14、16、18 of Wilson’s Disease Gene

Objectives To sequence and study the exon 8、12、14、16、18 of WD gene (hepatolenticular degeneration, Wilson's disease) and to establish the direct gene diagnosis. Methods Extract the genomic DNA from 102 WD patients and 82 normal controls, and amplify exon 8、12、14、16、18 of ATP7B gene by polymerase chain reaction(PCR). The amplification products of exon 8、exon 12 were digested with MspI and Tail respectively followed by sequencing the PCR products of exom 8、12、14、16、18 from all the patients and normal controls. The correlation between the mutation and clinical manifestation were stucdied. Results Digested by MspI through amelioration, 35 cases were abnormal in 102 patients. Sequence result shows that in the 102 patients 34.31% have homozygous or heterozygous Arg778Leu mutation in exon 8, 12 Leu770Leu polymorphism; and that for exon 12, 13 cases were abnormal using digestion by Tail, direct sequencing shows 12.75% of the cases have heterozygous Thr935Met mutation, one case was identified as heterozygous Arg919Gly, while Arg952Lys was detected in both patients and normal control. No mutation was found in exon14、16、18 of all volunteers. Conclusions Arg778Leu in exon 8 and Thr935Met in exon 12 are the hot point mutation of ATP7B gene, and PCR-MspI 、PCR-Tail digestion are reliable for molecular diagnosis in patients with Wilson's disease.