title: [Detection of mutant adenine phosphoribosyltransferase genes by polymerase chain reaction-single strand conformation polymorphism analysis].
abstract: Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis is a rapid and sensitive method to identify point mutations in a given sequence of genomic DNA. We tried to apply the PCR-SSCP to the diagnosis of adenine phosphoribosyltransferase (APRT) deficiency, which is an autosomal recessive hereditary disease leading to 2,8-dihydroxyadenine urolithiasis. Genomic APRT genes, with or without mutations, were amplified and labeled simultaneously with 32P-dCTP by PCR. When run in a 6% polyacrylamide gel containing 10% glycerol, two types of mutant genes, APRT*Q0 and APRT*J, gave bands clearly distinct from those of the respective normal APRT genes. Since heterozygotes as well as homozygotes for these mutant APRT genes can be detected in 2 days, PCR-SSCP should be a valuable method in the diagnosis of APRT deficiency and in screening a large population for APRT mutant genes.
authors.affiliation: Department of Urology, Faculty of Medicine, Kyoto University.
authors.forename: Y
authors.initials: Y
authors.lastname: Kaneko
authors.forename: H
authors.initials: H
authors.lastname: Takeuchi
authors.forename: J
authors.initials: J
authors.lastname: Takenawa
authors.forename: O
authors.initials: O
authors.lastname: Yoshida
authors.forename: S
authors.initials: S
authors.lastname: Takano
authors.forename: J
authors.initials: J
authors.lastname: Fujita
mesh_terms: D000228:Adenine Phosphoribosyltransferase / Q000235:genetics*; D005838:Genotype; D006801:Humans; D009154:Mutation; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic
chemical_list: D000228:Adenine Phosphoribosyltransferase