title: Multiplex primer extension reaction screening and oxidative challenge of glucose-6-phosphate dehydrogenase mutants in hemizygous and heterozygous subjects.
abstract: The primary objective of our study was to provide a simple and reliable assay for identifying the majority of G6PD genetic variants in the Chinese population. We optimized the multiplex primer extension reaction (MPER) assay for simultaneous screening of 14-point mutations in 98 G6PD-deficient subjects. Our data demonstrated that this method is precise, cost-effective and has successfully identified mutations in 97 out of 98 subjects, including all heterozygous mutants. We also detected a relatively high incidence (12.3%) of c.871G > A, and all of them harbored the silent mutation c.1311C > T. Apart from the screening program, the pharmacogenetic relationship between G6PD level and residual reduced glutathione (GSH) level was studied upon oxidative challenge by alpha-naphthol. The GSH levels were correlated with their status of G6PD deficiency, but no significant difference was observed between individual G6PD-deficient groups. Our data demonstrated the potentials of the MPER assay for characterization of G6PD deficiency and other genetic diseases.
authors.affiliation: Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, Hong Kong, SAR, China.
authors.forename: Chun Hay
authors.initials: CH
authors.lastname: Ko
authors.forename: Edmund
authors.initials: E
authors.lastname: Yung
authors.forename: Karen
authors.initials: K
authors.lastname: Li
authors.forename: Chung Leung
authors.initials: CL
authors.lastname: Li
authors.forename: Pak Cheung
authors.initials: PC
authors.lastname: Ng
authors.forename: Kwok Pui
authors.initials: KP
authors.lastname: Fung
authors.forename: Raymond Pui-On
authors.initials: RP
authors.lastname: Wong
authors.forename: Kit Man
authors.initials: KM
authors.lastname: Chui
authors.forename: Goldie Jia-Shi
authors.initials: GJ
authors.lastname: Gu
authors.forename: Tai Fai
authors.initials: TF
authors.lastname: Fok
mesh_terms: D002681:China / Q000453:epidemiology; D005820:Genetic Testing / Q000191:economics / Q000379:methods*; D005838:Genotype; D005954:Glucosephosphate Dehydrogenase / Q000032:analysis / Q000235:genetics*; D005955:Glucosephosphate Dehydrogenase Deficiency / Q000175:diagnosis / Q000235:genetics; D005978:Glutathione / Q000032:analysis; D006801:Humans; D015994:Incidence; D017720:Molecular Epidemiology; D009154:Mutation; D009284:Naphthols / Q000494:pharmacology; D021141:Nucleic Acid Amplification Techniques; D010084:Oxidation-Reduction
chemical_list: D009284:Naphthols; C029350:1-naphthol; D005954:Glucosephosphate Dehydrogenase; D005978:Glutathione