机构：[1]Department of Cell Biology and Medical Genetics, Kunming Medical University, Kunming, Yunnan Province, China[2]The First Affiliated Hospital, Kunming Medical University, Kunming, Yunnan Province, China昆明医科大学附属第一医院[3]Department of Histology and Embryology, Kunming Medical University, Kunming, Yunnan Province, China[4]Kunming City Maternal and Child Health Hospital, Kunming, Yunnan Province, China[5]Department of Pathogen Biology and Immunology, Kunming Medical University, Kunming, Yunnan Province, China[6]Department of Entomology, The Pennsylvania State University, University Park, Pennsylvania, USA
Introduction: 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. Conclusion: The SNaPshot method offers a specific and sensitive alternative for simultaneously interrogating multiple G6PD mutations.
基金：
National Natural Science
Foundation of China (# 31260264), High talent introduction
project of Yunnan province (# 2013HA026),
and National Institutes of Health, USA (U19AI0
89672).
第一作者机构：[1]Department of Cell Biology and Medical Genetics, Kunming Medical University, Kunming, Yunnan Province, China
共同第一作者：
通讯作者：
通讯机构：[*1]Department of Entomology, The Pennsylvania State University, 501 ASI Bldg., University Park, PA 16802, USA.[*2]Department of Cell Biology and Medical Genetics, Kunming Medical University, 1168 West Chunrong Road, Kunming, Yunnan Province 650500, China.
推荐引用方式(GB/T 7714)：
Zhang L.,Yang Y.,Liu R.,et al.A multiplex method for detection of glucose-6-phosphate dehydrogenase (G6PD) gene mutations[J].INTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY.2015,37(6):739-745.doi:10.1111/ijlh.12405.
APA：
Zhang, L.,Yang, Y.,Liu, R.,Li, Q.,Yang, F....&He, Y..(2015).A multiplex method for detection of glucose-6-phosphate dehydrogenase (G6PD) gene mutations.INTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY,37,(6)
MLA：
Zhang, L.,et al."A multiplex method for detection of glucose-6-phosphate dehydrogenase (G6PD) gene mutations".INTERNATIONAL JOURNAL OF LABORATORY HEMATOLOGY 37..6(2015):739-745
