title: A multiplex method for detection of glucose-6-phosphate dehydrogenase (G6PD) gene mutations.
abstract: BACKGROUND
  Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common human enzyme defect caused by G6PD gene mutations. This study aimed to develop a cost-effective, multiplex, genotyping method for detecting common mutations in the G6PD gene.
  
  
  METHODS
  We used a SNaPshot approach to genotype multiple G6PD mutations that are common to human populations in South-East Asia. This assay is based on multiplex PCR coupled with primer extension reactions. Different G6PD gene mutations were determined by peak retention time and colors of the primer extension products.
  
  
  RESULTS
  We designed PCR primers for multiplex amplification of the G6PD gene fragments and for primer extension reactions to genotype 11 G6PD mutations. DNA samples from a total of 120 unrelated G6PD-deficient individuals from the China-Myanmar border area were used to establish and validate this method. Direct sequencing of the PCR products demonstrated 100% concordance between the SNaPshot and the sequencing results.
  
  
  CONCLUSIONS
  The SNaPshot method offers a specific and sensitive alternative for simultaneously interrogating multiple G6PD mutations.
authors.affiliation: Department of Cell Biology and Medical Genetics, Kunming Medical University, Kunming, Yunnan Province, China.
authors.forename: L
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Cell Biology and Medical Genetics, Kunming Medical University, Kunming, Yunnan Province, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Yang
authors.affiliation: The First Affiliated Hospital, Kunming Medical University, Kunming, Yunnan Province, China.
authors.forename: R
authors.initials: R
authors.lastname: Liu
authors.affiliation: Department of Cell Biology and Medical Genetics, Kunming Medical University, Kunming, Yunnan Province, China.
authors.forename: Q
authors.initials: Q
authors.lastname: Li
authors.affiliation: Department of Cell Biology and Medical Genetics, Kunming Medical University, Kunming, Yunnan Province, China.
authors.forename: F
authors.initials: F
authors.lastname: Yang
authors.affiliation: Department of Histology and Embryology, Kunming Medical University, Kunming, Yunnan Province, China.
authors.forename: L
authors.initials: L
authors.lastname: Ma
authors.affiliation: The First Affiliated Hospital, Kunming Medical University, Kunming, Yunnan Province, China.
authors.forename: H
authors.initials: H
authors.lastname: Liu
authors.affiliation: Kunming City Maternal and Child Health Hospital, Kunming, Yunnan Province, China.
authors.forename: X
authors.initials: X
authors.lastname: Chen
authors.affiliation: Department of Pathogen Biology and Immunology, Kunming Medical University, Kunming, Yunnan Province, China.
authors.forename: Z
authors.initials: Z
authors.lastname: Yang
authors.affiliation: Department of Entomology, The Pennsylvania State University, University Park, Pennsylvania, USA.
authors.forename: L
authors.initials: L
authors.lastname: Cui
authors.affiliation: Department of Cell Biology and Medical Genetics, Kunming Medical University, Kunming, Yunnan Province, China.
authors.forename: Y
authors.initials: Y
authors.lastname: He
mesh_terms: D000293:Adolescent; D004252:DNA Mutational Analysis / Q000379:methods; D005260:Female; D060005:Genotyping Techniques; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics*; D005955:Glucosephosphate Dehydrogenase Deficiency / Q000175:diagnosis / Q000235:genetics; D006801:Humans; D008297:Male; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods; D009154:Mutation; D020641:Polymorphism, Single Nucleotide
chemical_list: D005954:Glucosephosphate Dehydrogenase
keywords: Glucose-6-phosphate dehydrogenase deficiency; SNaPshot assay; mutation detection