Abstract：Objective Since glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common hereditary hemolytic erythrocyte enzyme deficiency, most cases have single nucleotide mutations in the coding region, and current test methods for gene mutation have some missed detections, this study aimed to investigate the feasibility of RT-PCR sequencing in the detection of gene mutation in G6PD deficiency.Methods According to the G6PD/6GPD ratio, 195 children with anemia of unknown cause or who underwent physical examination between August 2013 and July 2014 were classified into G6PD-deficiency group with 130 children (G6PD/6GPD ratio Results In the G6PD-deficiency group, the detection rate of gene mutation was 100% and 13 missense mutations were detected, including one new mutation. In the control group, no missense mutation was detected in 28 boys; 13 heterozygous missense mutations, 1 homozygous same-sense mutation (C1191T) which had not been reported in China and abroad, and 14 single nucleotide polymorphisms of C1311T were detected in 37 girls. The control group showed a high rate of missed detection of G6PD deficiency (carriers) in the specimens from girls (35%, 13/37).Conclusions RTPCR sequencing has a high detection rate of G6PD gene mutation and a certain value in clinical diagnosis of G6PD deficiency.
LYU Rong-Yu,CHEN Xiao-Wen,ZHANG Min et al. Detection of gene mutation in glucose-6-phosphate dehydrogenase deficiency by RTPCR sequencing[J]. CJCP, 2016, 18(7): 630-634.
Minucci A, Moradkhani K, Hwang MJ, et al. Glucose-6- phosphate dehydrogenase(G6PD) mutations database: review of the "old" and update of the new mutations[J]. Blood Cells MolDis, 2012, 48(3): 154-165.
Liu WL, Li F, He ZX, et al. Glucose-6-phosphate dehydrogenase qingzhen: Identification of a novel splice mutation (IVS5-1 G>A)[J]. Pediatr Blood Cancer, 2012, 58(5): 825-826.
[7]
Jiang W, Yu G, Liu P, et al. Structure and function of glucose- 6-phosphate dehydrogenase-deficient variants in Chinese population[J]. Hum Genet, 2006, 119 (5): 463-478.
[8]
Yan JB, Xu HP, Xiong C, et al. Rapid and reliable detection of glucose-6-phosphate dehydrogenase (G6PD) gene mutations in Han Chinese using high-resolution melting analysis[J]. J Mol Diagn, 2010, 12(3): 305-311.
[9]
Tseng CP, Huang CL, Chong KY, et al. Rapid detection of glucose-6-phosphate dehydrogenase gene mutations by denaturing high-performance liquid chromatography[J]. Clin Biochem, 2005, 38(11): 973-980.
Myeroff LL, He H, Fink SP, et al. Mutation detection in the TGF-beta receptors and smad genes: RT-PCR and sequencing[J]. Methods Mol Biol, 2000, 142: 139-147.
[12]
Liu TC, Yen JS, Shen JS. Rapid molecular diagnosis of hemoglobin variants by RT-PCR of reticulocyte mRNA and direct sequencing[J]. Hemoglobin, 1992, 16(5): 379-388.
