title: Rapid detection of common Chinese glucose-6-phosphate dehydrogenase (G6PD) mutations by denaturing gradient gel electrophoresis (DGGE).
abstract: We describe here the use of denaturing gradient gel electrophoresis (DGGE) to detect the most common Chinese glucose-6-phosphate dehydrogenase (G6PD) variants, which are the single point mutations: G-->T at nt 1376, G-->A at 1388 both in exon 12 and A-->G at nt 95 in exon 02. In each case, the mutant allele resolves well from the normal allele(s). The distinct heteroduplex bands are characteristic of a particular genotype suggesting that this feature is very useful for identifying all heterozygous carriers for this and other X-linked diseases. When the analysis is extended to other exons, DGGE scans the gene and coupled with direct sequencing, it leads to the identification of new G6PD variation(s). With this approach, we identified a mutation in exon 9 which had not been reported in Hong Kong. Since DGGE can rapidly screen many unknown samples in one gel, this approach could be used to diagnose these G6PD mutations and to identify the at-risk for counselling.
authors: Department of Biochemistry, University of Hong Kong, Hong Kong.
authors: V M
authors: VM
authors: Lam
authors: W
authors: W
authors: Huang
authors: S T
authors: ST
authors: Lam
authors: C Y
authors: CY
authors: Yeung
authors: P H
authors: PH
authors: Johnson
mesh_terms: D000483:Alleles; D001483:Base Sequence; D002681:China; D004252:DNA Mutational Analysis / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005091:Exons; D005260:Female; D006580:Genetic Carrier Screening; D008040:Genetic Linkage; D014644:Genetic Variation; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics*; D005955:Glucosephosphate Dehydrogenase Deficiency / Q000201:enzymology* / Q000235:genetics*; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D017354:Point Mutation; D014960:X Chromosome / Q000235:genetics
chemical_list: D017931:DNA Primers; D005954:Glucosephosphate Dehydrogenase