Mutation Analysis Of PINK1 Gene By Using Denaturing High Performance Liquid Chromatography (DHPLC)

ObjectiveEstablish Phosphatase and tensinhomologue (PTEN)- induced kinase 1 (PINK1)gene diagnosis method by using denaturing high performance liquid chromatography(DHPLC). To detect the mutation of PINK1 gene in Chinese patients with early onset Parkinsonism (EOP) by using DHPLC.MethodsBy technolodgy of denaturing high performance liquid chromatography (DHPLC), We detect the mutation of PINK1 gene in 100 Chinese patients with EOP.The abnormal amplifications detected by DHPLC were sequenced. If we find the bas differentiation,we should apply the crosscheck analysis in 100 healthy person.ResultsAll the 8 coding exons of the PINK1 gene in 100 Chinese patients with EOP were detected, then we found 139 abnormal peak forms in the 900 amplicons,including the amplicons of exon2,4,5,7,8 of PINK1; abnormal peaks were detected in the amplicons of exon 8 in 63 persons,in the amplicons of exon 5 in 41 persons, in the amplicons of exon 2 of PINK1 gene in 22 persons, in the amlicons of exon 4 in 7 persons and in the amplicons of exon 7 in 6 persons.Further more, the amplicons with abnormal peaks were detected by DNA sequencing.We identified 1 novel single heterozygous missense mutation (C926G), 1 single heterozagous samesense mutation (C1362T); Novel (IVS4+18g→a) and known polymorphisms were identified, including of IVS2-7a→g,IVS5+68t→a,IVS4+18g→a,IVS4+72g→c,IVS5-5g→a,G1018A,A1562C,3'-UTR37a→t, 3'-UTR40g→a, and 5 amplicons with abnormal peaks were not identified the sequence change(false positive).ConclusionEstablish PINK1 gene diagnosis method by using denaturing high performance liquid chromatography(DHPLC). We identified 1 novelsingle heterozygous missense mutation (C926G), 1 single heterozagous samesense mutation (C1362T); Novel (IVS4+18g→a) and known polymorphisms were identified,including of IVS2-7a→g,IVS5+68t→a,IVS4+18g→a, IVS4+72g→c,IVS5-5g→a, G1018A,A1562C,3'-UTR37a→t, 3'-UTR40g→a...