title: Development of reverse dot-blot system for screening of mitochondrial DNA mutations associated with Leber hereditary optic atrophy.
abstract: We developed a diagnostic test based on the reverse dot-blot principle, in which five mitochondrial point mutations responsible for Leber hereditary optic neuropathy (LHON) were screened simultaneously. A series of wild-type and mutant oligonucleotides representing each mutation were covalently bound to a single nylon membrane strip. The target sites 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. In 11 cases (23%) the diagnosis of LHON could be confirmed (3460, 1; 9804, 1; 11778, 5; 14484, 3; 15257, 1). The results suggest that the clinical identification of the mitochondrial defect is not trivial and the availability of a rapid screening method simplifies the molecular analysis of these cases.
authors.affiliation: Center for Human Genetics, University of Leuven, Belgium.
authors.forename: E
authors.initials: E
authors.lastname: Schollen
authors.forename: P
authors.initials: P
authors.lastname: Vandenberk
authors.forename: J J
authors.initials: JJ
authors.lastname: Cassiman
authors.forename: G
authors.initials: G
authors.lastname: Matthijs
mesh_terms: D004252:DNA Mutational Analysis; D017931:DNA Primers; D004272:DNA, Mitochondrial / Q000235:genetics*; D006801:Humans; D008163:Luminescent Measurements; D009693:Nucleic Acid Hybridization; D015418:Optic Atrophies, Hereditary / Q000235:genetics*; D017354:Point Mutation; D016133:Polymerase Chain Reaction
chemical_list: D017931:DNA Primers; D004272:DNA, Mitochondrial