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Molecular Analysis of Glucose-6-Phosphate Dehydrogenase Gene Mutations in Bangladeshi Individuals

View Article: PubMed Central - PubMed

ABSTRACT

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.

No MeSH data available.


Related in: MedlinePlus

Proportion of G6PD enzyme activity levels for male and female participants compared to adjusted male median value.For each G6PD enzyme activity level (U/g Hb) shown in X-axis, the corresponding value in Y-axis indicates the number of participants. The numbers on top of each dotted line, shown as 10, 20, 30, and 60 on uppermost horizontal line of the graph indicate different cut-off values as percentages for the study population. 60% (shown as 60) of adjusted male median (shown as 100) is the upper limit of cut-off value and the participants with enzyme activities below 60% are considered deficient. Black portion of each bar indicates male participants, whereas gray portion of each bar indicates female participants.
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pone.0166977.g001: Proportion of G6PD enzyme activity levels for male and female participants compared to adjusted male median value.For each G6PD enzyme activity level (U/g Hb) shown in X-axis, the corresponding value in Y-axis indicates the number of participants. The numbers on top of each dotted line, shown as 10, 20, 30, and 60 on uppermost horizontal line of the graph indicate different cut-off values as percentages for the study population. 60% (shown as 60) of adjusted male median (shown as 100) is the upper limit of cut-off value and the participants with enzyme activities below 60% are considered deficient. Black portion of each bar indicates male participants, whereas gray portion of each bar indicates female participants.

Mentions: Table 2 illustrates G6PD activity profile for the study participants. A total of 121 clinically suspected participants, 79 males and 42 females were included in this study. The adjusted median G6PD enzyme activity for male participants was calculated by excluding five male participants who had enzyme activity less than 10% of the median value obtained for all male participants. The median G6PD activity for the study population was 12.73 U/g Hb (range: 0.7–24.57 U/g Hb). On the other hand, the adjusted male median enzyme activity (12.28 U/g Hb) was considered 100% enzyme activity (Fig 1) following Domingo et al. [17]. The Participants with enzyme activity less than 7.37 U/g Hb (60% of adjusted male median) were classified as deficient. Based on this cut-off value, 14 (12 males and 2 females) out of 121 participants were suspected as deficient for G6PD enzyme activity.


Molecular Analysis of Glucose-6-Phosphate Dehydrogenase Gene Mutations in Bangladeshi Individuals
Proportion of G6PD enzyme activity levels for male and female participants compared to adjusted male median value.For each G6PD enzyme activity level (U/g Hb) shown in X-axis, the corresponding value in Y-axis indicates the number of participants. The numbers on top of each dotted line, shown as 10, 20, 30, and 60 on uppermost horizontal line of the graph indicate different cut-off values as percentages for the study population. 60% (shown as 60) of adjusted male median (shown as 100) is the upper limit of cut-off value and the participants with enzyme activities below 60% are considered deficient. Black portion of each bar indicates male participants, whereas gray portion of each bar indicates female participants.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5120827&req=5

pone.0166977.g001: Proportion of G6PD enzyme activity levels for male and female participants compared to adjusted male median value.For each G6PD enzyme activity level (U/g Hb) shown in X-axis, the corresponding value in Y-axis indicates the number of participants. The numbers on top of each dotted line, shown as 10, 20, 30, and 60 on uppermost horizontal line of the graph indicate different cut-off values as percentages for the study population. 60% (shown as 60) of adjusted male median (shown as 100) is the upper limit of cut-off value and the participants with enzyme activities below 60% are considered deficient. Black portion of each bar indicates male participants, whereas gray portion of each bar indicates female participants.
Mentions: Table 2 illustrates G6PD activity profile for the study participants. A total of 121 clinically suspected participants, 79 males and 42 females were included in this study. The adjusted median G6PD enzyme activity for male participants was calculated by excluding five male participants who had enzyme activity less than 10% of the median value obtained for all male participants. The median G6PD activity for the study population was 12.73 U/g Hb (range: 0.7–24.57 U/g Hb). On the other hand, the adjusted male median enzyme activity (12.28 U/g Hb) was considered 100% enzyme activity (Fig 1) following Domingo et al. [17]. The Participants with enzyme activity less than 7.37 U/g Hb (60% of adjusted male median) were classified as deficient. Based on this cut-off value, 14 (12 males and 2 females) out of 121 participants were suspected as deficient for G6PD enzyme activity.

View Article: PubMed Central - PubMed

ABSTRACT

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.

No MeSH data available.


Related in: MedlinePlus