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A multiplex method for detection of glucose‐6‐phosphate dehydrogenase (G6 PD ) gene mutations
Author(s) -
Zhang L.,
Yang Y.,
Liu R.,
Li Q.,
Yang F.,
Ma L.,
Liu H.,
Chen X.,
Yang Z.,
Cui L.,
He Y.
Publication year - 2015
Publication title -
international journal of laboratory hematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.705
H-Index - 55
eISSN - 1751-553X
pISSN - 1751-5521
DOI - 10.1111/ijlh.12405
Subject(s) - multiplex , glucose 6 phosphate dehydrogenase , gene , microbiology and biotechnology , multiplex polymerase chain reaction , chemistry , dehydrogenase , biochemistry , genetics , biology , polymerase chain reaction , enzyme
Summary Introduction Glucose‐6‐phosphate dehydrogenase (G6 PD ) deficiency is the most common human enzyme defect caused by G6 PD gene mutations. This study aimed to develop a cost‐effective, multiplex, genotyping method for detecting common mutations in the G6 PD gene. Methods We used a SN aPshot approach to genotype multiple G6 PD mutations that are common to human populations in South‐East Asia. This assay is based on multiplex PCR coupled with primer extension reactions. Different G6 PD 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 G6 PD gene fragments and for primer extension reactions to genotype 11 G6 PD mutations. DNA samples from a total of 120 unrelated G6 PD ‐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 SN aPshot and the sequencing results. Conclusion The SN aPshot method offers a specific and sensitive alternative for simultaneously interrogating multiple G6 PD mutations.