title: Rapid detection of common mutation of arylsulfatase A in metachromatic leukodystrophy by polymerase chain reaction with a mismatched primer.
abstract: The most common mutation in late-onset metachromatic leukodystrophy is a cytosine-to-thymine substitution in exon VIII. This mutation caused a substitution of leucine for proline at amino acid residue 426. We developed a rapid and simple method for the detection of 426Pro-->Leu mutation by polymerase chain reaction with mismatched primer. Although the 426Pro-->Leu mutation does not alter recognition sequence for restriction enzymes, we created a Pst I restriction site using a 3'-primer mismatched at one nucleotide. As a result, the mutation can be detected as a Pst I restriction fragment length polymorphism.
authors.affiliation: Department of Inherited Metabolic Disease, National Institute of Neuroscience, NCNP, Tokyo, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Ohshima
authors.forename: M
authors.initials: M
authors.lastname: Sasaki
authors.forename: J
authors.initials: J
authors.lastname: Takahashi
authors.forename: N
authors.initials: N
authors.lastname: Sakuragawa
mesh_terms: D017668:Age of Onset; D002553:Cerebroside-Sulfatase / Q000172:deficiency / Q000235:genetics*; D002869:Chromosome Aberrations; D025063:Chromosome Disorders; D004252:DNA Mutational Analysis; D017931:DNA Primers / Q000302:isolation & purification*; D005260:Female; D006801:Humans; D007966:Leukodystrophy, Metachromatic / Q000175:diagnosis* / Q000235:genetics; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D017931:DNA Primers; D002553:Cerebroside-Sulfatase