Direct sequencing and analysis of exon 8,12,18 of ATP7B gene of Wilson′s disease with mutations

Objective To make PCR and sequencing the exon 8,12,18 of the ATP7B gene in Wilson′disease(WD) patients,and to study the frequency of the mutations.Methods The genomic DNA from 30 WD patients and 10 normal controls was extracted,and the relative parts of exon 8,12,18 of ATP7B gene were amplified by PCR,which were directly sequenced and analyzed.Results The normal controls were all normol.The exon 8 with mutations was found in the 12 WD patients with the mutation ratio of 40.0%.The sequencing results showed that the WD patients had homozygous or heterozygous Arg778leu mutation in exon 8.The exon 12 with 6 mutations was found in 4 WD patients,and 2 WD patients had double mutations.No mutations were found in exon 18.ConclusionExon 8 and exon 12 are the mutation hot points of ATP7B gene,but exon 18 is not.