Multiplex primer extension reaction screening and oxidative challenge of glucose-6-phosphate dehydrogenase mutants in hemizygous and heterozygous subjects.
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.
Ko
Chun Hay
CH
Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, Hong Kong, SAR, China.
Yung
Edmund
E
Li
Karen
K
Li
Chung Leung
CL
Ng
Pak Cheung
PC
Fung
Kwok Pui
KP
Wong
Raymond Pui-On
RP
Chui
Kit Man
KM
Gu
Goldie Jia-Shi
GJ
Fok
Tai Fai
TF
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
D009284:Naphthols; C029350:1-naphthol; D005954:Glucosephosphate Dehydrogenase; D005978:Glutathione