title: A comprehensive method to scan for point mutations of the glucose 6 phosphate dehydrogenase gene.
abstract: We have developed a fast and comprehensive method to scan for point mutations in a gene on X chromosome. A target region of the gene is first amplified. Then, using the amplified product as a template, PCR is carried out with multiple short-length forward primers arrayed in tandem in the scanned region, and a common reverse primer. The absence of amplified product defines the site of a mutation within a narrow region of the primer recognition site. To evaluate our method, point mutations in exon 12 of the human glucose-6-phosphate dehydrogenase (G6PD) gene were used as a model system. Out of 12 Singaporean G6PD-deficient patients, 6 cases were shown by the method to have a nucleotide change in this exon. Sequence analysis confirmed the presence of a nucleotide change in the region identified by our scanning. Thus, our method is accurate in localizing mutations within a narrow region, and allows large numbers of samples to be handled simultaneously.
authors.affiliation: Faculty of Health Science, Kobe University School of Medicine, Tomogaoka, Suma-ku, Kobe 654-01, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Shirakawa
authors.forename: K
authors.initials: K
authors.lastname: Nishiyama
authors.forename: P S
authors.initials: PS
authors.lastname: Lai
authors.forename: M
authors.initials: M
authors.lastname: Matsuo
mesh_terms: D041321:Chromosomes, Human, X; D004252:DNA Mutational Analysis / Q000379:methods*; D005069:Evaluation Studies as Topic; D005091:Exons; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics*; D005955:Glucosephosphate Dehydrogenase Deficiency / Q000175:diagnosis; D006801:Humans; D007231:Infant, Newborn; D008297:Male; D016297:Mutagenesis, Site-Directed; D017354:Point Mutation / Q000235:genetics*
chemical_list: D005954:Glucosephosphate Dehydrogenase