Sarker Suprovath Kumar, Islam Md Tarikul, Eckhoff Grace, Hossain Mohammad Amir, Qadri Syeda Kashfi, Muraduzzaman A K M, Bhuyan Golam Sarower, Shahidullah Mohammod, Mannan Mohammad Abdul, Tahura Sarabon, Hussain Manzoor, Akhter Shahida, Nahar Nazmun, Shirin Tahmina, Qadri Firdausi, Mannoor Kaiissar
Laboratory of Genetics and Genomics, Institute for Developing Science and Health Initiatives, Mohakhali, Dhaka, Bangladesh.
Department of Paediatric Medicine, KK Women's and Children's Hospital, 100 Bukit Timah Road, Singapore, Singapore.
PLoS One. 2016 Nov 23;11(11):e0166977. doi: 10.1371/journal.pone.0166977. eCollection 2016.
Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common X-linked human enzyme defect of red blood cells (RBCs). Individuals with this gene defect appear normal until exposed to oxidative stress which induces hemolysis. Consumption of certain foods such as fava beans, legumes; infection with bacteria or virus; and use of certain drugs such as primaquine, sulfa drugs etc. may result in lysis of RBCs in G6PD deficient individuals. The genetic defect that causes G6PD deficiency has been identified mostly as single base missense mutations. One hundred and sixty G6PD gene mutations, which lead to amino acid substitutions, have been described worldwide. The purpose of this study was to detect G6PD gene mutations in hospital-based settings in the local population of Dhaka city, Bangladesh. Qualitative fluorescent spot test and quantitative enzyme activity measurement using RANDOX G6PDH kit were performed for analysis of blood specimens and detection of G6PD-deficient participants. For G6PD-deficient samples, PCR was done with six sets of primers specific for G6PD gene. Automated Sanger sequencing of the PCR products was performed to identify the mutations in the gene. Based on fluorescence spot test and quantitative enzyme assay followed by G6PD gene sequencing, 12 specimens (11 males and one female) among 121 clinically suspected patient-specimens were found to be deficient, suggesting a frequency of 9.9% G6PD deficiency. Sequencing of the G6PD-deficient samples revealed c.C131G substitution (exon-3: Ala44Gly) in six samples, c.G487A substitution (exon-6:Gly163Ser) in five samples and c.G949A substitution (exon-9: Glu317Lys) of coding sequence in one sample. These mutations either affect NADP binding or disrupt protein structure. From the study it appears that Ala44Gly and Gly163Ser are the most common G6PD mutations in Dhaka, Bangladesh. This is the first study of G6PD mutations in Bangladesh.
葡萄糖-6-磷酸脱氢酶(G6PD)缺乏症是一种常见的、与X染色体相关的人类红细胞酶缺陷疾病。具有这种基因缺陷的个体在暴露于诱导溶血的氧化应激之前看起来是正常的。食用某些食物,如蚕豆、豆类;感染细菌或病毒;以及使用某些药物,如伯氨喹、磺胺类药物等,可能会导致G6PD缺乏个体的红细胞溶解。导致G6PD缺乏症的基因缺陷大多已被确定为单碱基错义突变。全球已描述了160种导致氨基酸替代的G6PD基因突变。本研究的目的是在孟加拉国达卡市当地人群的医院环境中检测G6PD基因突变。使用兰多克斯G6PDH试剂盒进行定性荧光斑点试验和定量酶活性测量,以分析血样并检测G6PD缺乏的参与者。对于G6PD缺乏的样本,使用六组针对G6PD基因的引物进行聚合酶链反应(PCR)。对PCR产物进行自动桑格测序以鉴定基因中的突变。基于荧光斑点试验、定量酶测定以及随后的G6PD基因测序,在121份临床疑似患者样本中发现12份样本(11名男性和1名女性)存在缺陷,表明G6PD缺乏症的发生率为9.9%。对G6PD缺乏样本的测序揭示,六个样本中存在c.C131G替代(外显子3:Ala44Gly),五个样本中存在c.G487A替代(外显子6:Gly163Ser),一个样本的编码序列中存在c.G949A替代(外显子9:Glu317Lys)。这些突变要么影响烟酰胺腺嘌呤二核苷酸磷酸(NADP)结合,要么破坏蛋白质结构。从该研究看来,Ala44Gly和Gly163Ser是孟加拉国达卡最常见的G6PD突变。这是孟加拉国关于G6PD突变的首次研究。