Rapid detection of common mutation of arylsulfatase A in metachromatic leukodystrophy by polymerase chain reaction with a mismatched primer.
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.
Ohshima
T
T
Department of Inherited Metabolic Disease, National Institute of Neuroscience, NCNP, Tokyo, Japan.
Sasaki
M
M
Takahashi
J
J
Sakuragawa
N
N
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*
D017931:DNA Primers; D002553:Cerebroside-Sulfatase