DEVELOPMENT OF REVERSE DOT-BLOT SYSTEM FOR SCREENING OF MITOCHONDRIAL-DNA MUTATIONS ASSOCIATED WITH LEBER HEREDITARY OPTIC ATROPHY

We developed a diagnostic test based on the reverse dot-blot principle , in which five mitochondrial point mutations responsible for Leber he reditary optic neuropathy (LHON) were screened simultaneously. A serie s of wild-type and mutant oligonucleotides representing each mutation were covalently bound to a single nylon membrane strip. The target sit es were amplified in a multiplex PCR and the products were hybridized to the membrane. Detection is based on chemiluminescence. To test the developed assay, 47 patients suspected of having LHON were screened. I n 11 cases (23%) the diagnosis of LHON could be confirmed (3460, 1; 98 04, 1; 11778, 5; 14484, 3; 15257, 1). The results suggest that the cli nical identification of the mitochondrial defect is not trivial and th e availability of a rapid screening method simplifies the molecular an alysis of these cases.