Point mutations in mitochondrial DNA (mtDNA) frequently cause the occurrence or the loss of a recognition site for the restriction endonuclease, and their diagnosis is often based on analysis of restriction fragments length polymorfism (RFLP). In the case of incomplete DNA digestion, RFLP analysis can give false negative results (when a mutation produces new restriction site) or false positive results (when a mutation abolishes the restriction site). The majority of cases of Leber’s hereditary optic neuropathy (LHON) are caused by mtDNA point mutations G3460A and G11778A. We have used the method of multiplex allele-specific PCR for diagnosis of these two LHON mutations. This assay is based on specific and simultaneous amplification of mutated mtDNA fragments and it is also faster and less expensive than RFLP analysis.

        Key words: Leber’s hereditary optic neuropathy (LHON), allele-specific PCR, mtDNA point mutations.