Application of Denaturing High-performance Liquid Chromatography for Detecting Mutations of Mitochondrial DNA

Objective To establish and evaluate denaturing high-performance liquid chromatography(DHPLC)as a rapid and efficient technique of detecting mutations of mitochondrion gene.Methods Collected four mitochondrial myopathy patients and 50 healthy individuals as control.The genome DNA was extracted.The whole tRNA gene of mitochondria was amplified by polymerase chain reaction(PCR).The PCR products were analyzed by denaturing high-performance liquid chromatography(DHPLC).The samples with abnormal profiles were sequenced.Results The examinations revealed that patient l had a homoplasmic A1625G mutation in tRNA-Val gene,patient 2 had a heteroplasmic A1625G/A mutation in tRNA-Val gene,patient 3 had a heteroplasmic A10411C/A mutation in tRNA-Arg gene and patient 4 had a homoplasmic T5553C mutation in tRNATrp gene.Conclusion The DHPLC technique provides an efficient,sensitive and rapid method for detecting mutations of mitochondrial DNA.