[Polymerase chain reaction with subsequent direct gene sequence analysis. A possibility for molecular analysis in...

Molecular analyses are of increasing importance in clinical research as well as for diagnostic evaluations. Gene alterations are involved in the pathogenesis of numerous diseases and are described in an increasing number of conditions. A method is presented which allows for rapid and detailed nucleotide sequence analysis of a segment of a gene. Buccal epithelial cells of a patient are obtained by means of a mouth wash with 10 ml water. DNA molecules are amplified in vitro using the polymerase chain reaction, followed by direct sequence analysis of the product. Using this technique, a sequence analysis of a gene segment is possible within 1-2 days. As an example, the analysis of a segment of the gene for uroporphyrinogen decarboxylase of patients with cutanea tarda is described.