Detect MFN2 Gene Mutation By Using Denaturing High Performance Liquid Chromatography

ObjectiveEstablish MFN2 gene diagnosis method by using denaturing high performance liquid chromatography(DHPLC). To detect the mutation of mitochondrial GTPase mitofusin 2, (MFN2) gene in 45 Chinese patients with Charcot-Marie-Tooth disease (CMT) by using denaturing high performance liquid chromatography (DHPLC).MethodsBy technolodgy of denaturing high performance liquid chromatography(DHPLC), We detect the mutation of mitochondrial GTPase mitofusin 2, (MFN2)gene in 45 Chinese patients with Charcot-Marie-Tooth disease(CMT). The abnormal amplifications detected by DHPLC were sequenced. If we find the bas differentiation, we we should apply the cross-check analysis in 100 healthy person. and Segregation analysis in family to make sure that whether it is a polymorphism or a mutation.ResultsAll the 17 coding exons of the MFN2 gene in 45 Chinese patients with Charcot-Marie-Tooth disease were detected, then we found 9 abnormal peak forms in 9 CMT patients. There is one in exon 2 and eight in exon 3; totally there are three kinds of abnormal peak forms, one in exon 2 and two in exon 3.Further more, the amplifications with abnormal peak form were sequenced. We found three heterozygous mutations which were 281G→A, 395 G→A and 408A→T. We found a pathogenic mutant 281G→A (Arg94Gln) which was previously identified, a novel nonsense mutation 408A→T(Val136Val)and a novel missense mutation 395 G→A (Cys132Tyr)ConclusionEstablish MFN2 gene diagnosis method by using denaturing high performance liquid chromatography(DHPLC). At the first time, we detected the MFN2 gene mutations using denaturing high performance liquid chromatography. In our CMT group, we found that a pathogenic mutant 281G→A (Arg94Gln), which was previously identified, a novel nonsense mutation 408A→T(Val) and a novel missense mutation 395 G→A (Cys132Tyr)...