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RESEARCH ARTICLE |
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Year : 2022 | Volume
: 10
| Issue : 3 | Page : 98-103 |
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Glucose-6-phosphate dehydrogenase deficiency and its association with malaria in the administrative divisions of Lagos State, Nigeria
Oladayo Musa Babalola1, Adetunji Alabi Alli1, Mojeed Ayoola Ashiru2
1 Department of Biochemistry, Faculty of Basic Medical Sciences, College of Medicine, University of Lagos, Lagos, Nigeria 2 Department of Chemical Sciences, Biochemistry Unit, College of Pure and Applied Sciences, Fountain University, Osogbo, Nigeria
Date of Submission | 20-Oct-2022 |
Date of Decision | 23-Oct-2022 |
Date of Acceptance | 14-Nov-2022 |
Date of Web Publication | 05-Dec-2022 |
Correspondence Address: Mr. Adetunji Alabi Alli Department of Biochemistry, Faculty of Basic Medical Sciences, College of Medicine, University of Lagos, Lagos Nigeria
 Source of Support: None, Conflict of Interest: None  | Check |
DOI: 10.4103/njecp.njecp_14_22
Background: Glucose-6-phosphate dehydrogenase (G6PD) deficiency is one of red blood cells' most common inherited enzyme disorders. It is currently believed to affect approximately 500 million individuals worldwide. The deficiency could result in several hematological conditions, including acute hemolytic anemia, neonatal jaundice, and kernicterus. Aim: This study aimed to determine the prevalence of G6PD deficiency in the five administrative divisions of Lagos State. The association between malaria and G6PD deficiency was also investigated. Materials and Methods: A total of 105 participants, comprising 63 (60%) males and 42 (40%) females, were recruited from five locations for this study. Two milliliters of venous blood were collected and divided into three portions for G6PD enzyme assay, hematological parameter, and malaria diagnosis. G6PD enzyme level was determined using a quantitative spectrophotometric assay, whereas the malaria parasite was examined using microscopy and rapid diagnostic test kits. Results: An overall prevalence of G6PD deficiency was 21%. There was no significant difference in prevalence between males (22.2%) and females (19%), whereas a marginally higher G6PD activity in males (10.15 ± 0.50 vs. 8.61 ± 0.31 U/g Hb) (P < 0.05) was recorded. Although there were slight differences in prevalence obtained in the five sampling locations, however, the one-way analysis of variance of the G6PD activity levels showed no significant difference between any pairs (P = 0.05). Furthermore, the results obtained from this study showed no association between malaria and G6PD deficiency (χ2 = 1.432, P = 0.231). Conclusions: The study found a relatively high prevalence of G6PD deficiency in the Nigerian subpopulation, indicating that G6PD deficiency is common in this environment. This emphasizes the need for a quantitative G6PD assay as part of laboratory investigations for those presenting with an episode of acute hemolytic anemia in this geographical region of the country.
Keywords: Anemia, glucose-6-phosphate dehydrogenase, Lagos, malaria, spectrophotometry
How to cite this article: Babalola OM, Alli AA, Ashiru MA. Glucose-6-phosphate dehydrogenase deficiency and its association with malaria in the administrative divisions of Lagos State, Nigeria. Niger J Exp Clin Biosci 2022;10:98-103 |
How to cite this URL: Babalola OM, Alli AA, Ashiru MA. Glucose-6-phosphate dehydrogenase deficiency and its association with malaria in the administrative divisions of Lagos State, Nigeria. Niger J Exp Clin Biosci [serial online] 2022 [cited 2023 Feb 9];10:98-103. Available from: https://www.njecbonline.org/text.asp?2022/10/3/98/362648 |
Introduction | |  |
Glucose-6-phosphate dehydrogenase (G6PD) deficiency is one of red blood cells' (RBCs) most common inherited enzyme disorders.[1] It is currently believed to affect approximately 500 million individuals worldwide.[2] It is an X-linked recessive genetic condition caused by mutations in the G6PD gene, which causes low or absent enzyme expression and a corresponding deficit.[3] It is believed to be more common in males than in females.
G6PD is a cytoplasmic enzyme in all life forms.[4] It is the rate-limiting enzyme in the pentose phosphate pathway (PPP), which converts glucose to a five-carbon pentose sugar needed for nucleic acid biosynthesis.[5],[6] This enzyme also converts oxidized nicotinamide adenine dinucleotide phosphate (NADP+) to its reduced form (NADPH).[7] NADPH maintains intracellular homeostasis of the reduced glutathione, which is directly involved in the preservation of RBC membrane and hemoglobin from the harmful effects of reactive oxygen species.[8],[9]
G6PD is found in all cells; however, its absence has substantial hematological consequences, with hemolytic anemia and hyperbilirubinemia being the most common clinical symptoms. This is because erythrocytes lack mitochondria and some other organelles. Thus, PPP is the sole source of NADPH. Therefore, RBCs depend on G6PD activity to generate NADPH for protection against oxidative damage.[10]
In most cases, people with G6PD deficiency are asymptomatic.[11] Exposure to exogenous substances, such as certain foods, particularly those from the leguminous family, infections, and some conventional antimalaria drugs, can cause acute hemolytic crises, leading to life-threatening anemia.[12]
G6PD deficiency has a wide geographical distribution, ranging from very rare among indigenous populations of Northern Europe to over 20% in portions of Southern Europe and Asia and up to 40% in some areas of Southern East Asia, Africa, and the Middle East.[13],[14] Worldwide, high frequencies occur in areas where malaria has been endemic. As a result, it has been claimed that G6PD deficiency may provide natural protection from malaria infection.[5],[11],[15]
Studies have reported various rates of G6PD deficiency in some parts of Nigeria. For instance, 14.4% has been reported in Sokoto, North West Nigeria.[16] Egesie et al.[17] have also reported a 20% prevalence in Jos, North Central, Nigeria. The World Health Organization has recommended a routine national screening program in regions where the prevalence is 3% and above.[18] However, this has not yet become a practice in many parts of the country. Therefore, the present study aimed to determine the prevalence of G6PD deficiency in Lagos, Nigeria's largest city, commercial nerve center, and former administrative headquarters. A representative sampling of the state was done using her five administrative divisions. Of the several methods available for G6PD testing, the quantitative assay is said to be the gold standard. Therefore, this study employed quantitative spectrophotometric analysis to determine the prevalence of G6PD deficiency in the five administrative divisions of Lagos state. We also investigated the relationship between malaria infection and G6PD deficiency.
Materials and Methods | |  |
Study site
This cross-sectional study occurred between August and October 2021 in the five administrative divisions of Lagos State. Lagos State is located in Nigeria's southwestern geopolitical zone. The state is perhaps the most economically important in Nigeria, as it contains Lagos, the country's largest city. It is divided into five administrative divisions, namely, Ikeja, Badagry, Ikorodu, Lagos (Eko), and Epe, which are further divided into 20 local government area[19], as shown in [Figure 1]. The state is essentially a Yoruba society inhabited by its subnationality of Ogus in Badagry and Remos and Ijebus are found in Ikorodu and Epe, whereas Aworis form the indigenous population of Lagos and Ikeja divisions.
Sample collection
The blood sample was collected from a convenient peripheral vein under aseptic conditions. Two milliliters (2 mL) of venous blood were collected using a 5 mL syringe. The blood was introduced into a labeled bottle containing dipotassium salt of ethylenediaminetetraacetic acid for hematological parameter measurement, G6PD enzyme assay, and malaria diagnosis
Glucose-6-phosphate dehydrogenase activity determination
G6PD kit obtained from Fortress diagnostics (with product code: BXC0571) was used. The kit functions on the principle of reaction described by Beutler,[20] in which NADPH is produced from NADP+. The NADPH produced is measured spectrophotometrically at 340 nm.
The assay was performed according to the manufacturer's instructions. Briefly, 0.2 mL of whole blood was washed with 2 mL aliquot of 0.9% NaCl solution and then centrifuged for 10 min at 3,000 rpm. This process was repeated three times. The washed erythrocytes were then suspended in 0.5 mL of digitonin solution and allowed to stand for 15 min at 4°C, followed by centrifugation. The supernatant was incubated with substrate and cofactor at 37°C for 10 min in an electric oven. This was then used in ultraviolet/visible spectrophotometer for the enzyme assay within 2 h of preparation.
Automated techniques were used to determine the RBC count and hemoglobin content of the samples. The Sysmex percentage absolute count and Sysmex CyFlow Counter were used.
The G6PD activity was expressed in international units per gram of hemoglobin (IU/g Hb) using the following calculation:

Values ≤6.90 U/g Hb were considered deficient
Values >6.97 were considered sufficient (normal)
Microscopic diagnosis
Malaria diagnosis was conducted using two methods: examining the Giemsa stained thick and thin blood smears using a light microscope and detecting malaria parasite antigen using a rapid diagnostic test (RDT) kit.
Microscopic examination of malaria parasites
Eight microliters of blood samples were placed on labeled slides using a micropipette to make thick films. Each sample was spread with the beveled edge of the spreader until the entire circle of 12-mm diameter was covered evenly. Films were dried on a dust-free surface and fixed in absolute ethanol. The films were allowed to dry again and then stained using Giemsa stain. The stained films were examined under a light microscope by a WHO-certified malariologist.
Rapid diagnostic test diagnosis of malaria
The RDT kit obtained from CareStart™ Malaria pf (HRP2) Ag was used. The test was carried out according to the manufacturer's instructions. Briefly, 5 μL of blood sample collected using a specimen transfer device provided was added into the “S” well of the cassette in the test kit. Two drops (60 μL) of the buffer solution were added into the “A” well, and the result was read after 20 min.
Statistical analysis
All data were analyzed using GraphPad Prism software version 9 of (GraphPad Software Inc, California, USA). Indices of central statistical location, mean median, mode, and standard deviation were used to describe continuous data. The one-way analysis of variance was used to assess the significance among the means of more than two groups. The Student's t-test was used for two groups, whereas Chi-square was used to compare categorical data. A P < 0.05 was considered statistically significant. The results are reported in tables.
Results | |  |
Population distribution of the studied participants
A total of 105 participants, comprising 63 (60%) males and 42 (40%) females, were studied. Of this studied population, participants from the Ikeja administrative division made up the most significant percentage, 30. Participants from the Ikorodu division were the second largest, with 23, followed by 21 from Badagry, 17 from Lagos (Eko), and 14 from the Epe division. Participants ranged in age from 12 to 55 years. The mean age was 28.7 years (±0.92 SEM). Participants between the ages of 21–30 years and 31–40 years made up the most significant groups, with 48.8% and 21%, respectively. [Table 1] shows the population distribution.
G6PD activity levels of the studied participants
The G6PD activity of the studied population ranged from 1.01 to 16.81 U/g Hb, with a mean activity of 9.51 ± 3.40 U/g Hb. The overall prevalence of G6PD deficiency was 21% in all studied populations –22 (21%) of the participants were G6PD deficient, comprising 14 males with a prevalence of 22.2% and eight females with a prevalence of 19% (male: female ratio of 1.2:1). The remaining 83 (79%) of the participants were G6PD normal, comprising 49 males (77.8%) and 34 females (81%).
When the G6PD activities of males were compared to those of females (10.15 ± 0.50 vs. 8.61 ± 0.31), there was a statistically significant difference between the mean G6PD activity levels (P < 0.05), as shown in [Table 2]. | Table 2: Glucose-6-phosphate dehydrogenase activity levels and sex of all participants
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Distribution and comparison of glucose-6-phosphate dehydrogenase activity levels in the five sampling locations
When G6PD activities obtained from participants in the five different locations were compared with the standard value, Ikeja participants had a slightly higher prevalence (23.3%) than Lagos (20%), Badagry (19.1%), Ikorodu (17.4%), and Epe (7.1%) as shown in [Table 3]. However, Turkey's multiple comparisons of G6PD activity levels showed no significant difference between any pairs (P = 0.05). [Table 3] compares the G6PD activity level among the participants in the five different locations. | Table 3: Distribution and comparison of glucose-6-phosphate dehydrogenase activity levels in the five different sampling locations
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Association of glucose-6-phosphate dehydrogenase deficiency and malaria
Eight of 105 studied participants, consisting of five G6PD normal and three G6PD deficient participants, were malaria positive, whereas the remaining 78 G6PD normal and 19 G6PD deficient participants were malaria negative. When compared using the Chi-square test, the results showed no significant difference between G6PD normal and G6PD deficient participants concerning malaria (χ2 = 1.432, P = 0.231). [Table 4] shows the comparison. | Table 4: Association of glucose-6-phosphate dehydrogenase deficiency and malaria
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Discussion | |  |
G6PD is a PPP enzyme whose activity produces NADPH, which is needed to protect RBCs from oxidative stress caused by pro-oxidants such as certain foods, infections, and traditional antimalarial drugs.[21] The deficiency of this enzyme could result in several hematological conditions, including acute hemolytic anemia, neonatal jaundice, and kernicterus.[22],[23] In this study, we investigated the prevalence of G6PD deficiency in Lagos state. Several studies have reported differing prevalent values in different parts of Nigeria. However, no such study had been reported in Lagos, Africa's largest economy and a mega city housing virtually all the ethnic groups in Nigeria.
The overall prevalence obtained in this study using the quantitative spectrophotometric assay was 21%. There was no significant difference in prevalence between males (22.2%) and females (19%), whereas a marginally higher G6PD activity was recorded in males. These results are consistent with those of Egesie et al.,[17] as well as the results of Obasa et al.,[24] who found a prevalence of a 1.1:1 male-to-female ratio. In addition, there were no intra-ethnic differences in the prevalence recorded, suggesting the possibility of having only one G6PD variant in this region of the country. Meanwhile, this study recorded a higher prevalence than the 14.4% previously reported in the northern part of the country.[16] This could be due to geographic location or ethnic differences. It has been reported that the distribution and frequency of G6PD deficiency are related to specific geographic locations and are associated with different ethnic groups.[4] Various prevalence values have been reported in the different regions of Africa. For instance, 22.5% has been reported in Congo (Brazzaville), 15.7% in Mali, 13.0% in Uganda, and 9%–15.5% in Gabon.[25],[26] Surprisingly, the value obtained in this study was also higher than the 15.4% earlier reported in the same region.[27] This could be accounted for by intra-ethnic differences in the expression and distribution of G6PD variants, as suggested by Sathupak et al.[28] in their report. However, it could also be attributed to the method used in the investigation. Studies have shown that fluorescence spot tests may give false-negative results in heterozygote females and in homozygote males who have experienced a recent episode of hemolysis.[29] This suggests that the quantitative assay used in the present study could have possibly been able to diagnose many heterozygous females who may have been missed by the fluorescence spot test used in the previous study. Thus, the quantitative assay tends to give a higher prevalence than other methods. It is equally important to mention that the value obtained in the present study was significantly lower than the 39.2% reported by Amiwero and Olatunji,[30] 37.3% by Obasa et al.,[24] and 52.2% by George et al.[31] among jaundiced children. These previous studies, however, enrolled a different target population.
There have been conflicting reports on the association between G6PD deficiency and malaria. Several studies have suggested a protective effect,[32],[33] whereas another study by Mbanefo et al.[34] found no association. Although our results agreed with Mbanefo et al., however, a conclusive inference could not be drawn due to the few malaria patients enrolled in this study.
One of the significant limitations of this study was our inability to carry out molecular analysis of the samples to identify specific variants of G6PD prevalent in this location. Furthermore, financial constraints and the unavailability of other resources limited the study's sample size. Further research focusing on identifying the prevalent variants, and more extensive population studies at the national level should be carried out.
Conclusions | |  |
This study discovered a 21% prevalence of G6PD deficiency in the Lagos State administrative divisions (Ikeja, Badagry, Ikorodu, Lagos, and Epe) using the quantitative spectrophotometric assay. The mean G6PD activity was significantly higher in males compared to females, and more male than female participants were G6PD deficient. There was no statistically significant difference in the prevalence when the five locations were compared.
In conclusion, this study found a relatively high prevalence of G6PD deficiency in the Nigerian subpopulation, indicating that G6PD deficiency is common in this environment. This emphasizes the need for a quantitative G6PD assay as part of laboratory investigations for those presenting with an episode of acute hemolytic anemia in this country's geographical region.
Acknowledgment
The authors are grateful to the entire staff of the Department of Biochemistry, Nigeria Institute of Medical Research (NIMR), especially Dr. Ajibaye and Mr. Christopher, for their help in the microscopic viewing of the malaria parasite. The authors also thank Mr. Tajudeen of the Department of Haematology for his numerous contributions to hematological analysis.
Financial support and sponsorship
Nil.
Conflicts of interest
There are no conflicts of interest.
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[Figure 1]
[Table 1], [Table 2], [Table 3], [Table 4]
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