Diagnosis Of Glucose-6-phosphate Dehydrogenase Deficiency Research Articles

Field assessment of the operating procedures of a semi-quantitative G6PD Biosensor to improve repeatability of routine testing.

In remote communities, diagnosis of G6PD deficiency is challenging. We assessed the impact of modified test procedures and delayed testing for the point-of-care diagnostic STANDARD G6PD (SDBiosensor, RoK), and evaluated recommended cut-offs. We tested capillary blood from fingerpricks (Standard Method) and a microtainer (BD, USA; Method 1), venous blood from a vacutainer (BD, USA; Method 2), varied sample application methods (Methods 3), and used micropipettes rather than the test's single-use pipette (Method 4). Repeatability was assessed by comparing median differences between paired measurements. All methods were tested 20 times under laboratory conditions on three volunteers. The Standard Method and the method with best repeatability were tested in Indonesia and Nepal. In Indonesia 60 participants were tested in duplicate by both methods, in Nepal 120 participants were tested in duplicate by either method. The adjusted male median (AMM) of the Biosensor Standard Method readings was defined as 100% activity. In Indonesia, the difference between paired readings of the Standard and modified methods was compared to assess the impact of delayed testing. In the pilot study repeatability didn't differ significantly (p = 0.381); Method 3 showed lowest variability. One Nepalese participant had <30% activity, one Indonesian and 10 Nepalese participants had intermediate activity (≥30% to <70% activity). Repeatability didn't differ significantly in Indonesia (Standard: 0.2U/gHb [IQR: 0.1-0.4]; Method 3: 0.3U/gHb [IQR: 0.1-0.5]; p = 0.425) or Nepal (Standard: 0.4U/gHb [IQR: 0.2-0.6]; Method 3: 0.3U/gHb [IQR: 0.1-0.6]; p = 0.330). Median G6PD measurements by Method 3 were 0.4U/gHb (IQR: -0.2 to 0.7, p = 0.005) higher after a 5-hour delay compared to the Standard Method. The definition of 100% activity by the Standard Method matched the manufacturer-recommended cut-off for 70% activity. We couldn't improve repeatability. Delays of up to 5 hours didn't result in a clinically relevant difference in measured G6PD activity. The manufacturer's recommended cut-off for intermediate deficiency is conservative.

Secondary Reporting of G6PD Deficiency on Newborn Screening

In April 2019, the Alberta Newborn Screening Program expanded to include screening for classic galactosemia using a two-tier screening approach. This approach secondarily identifies infants with glucose-6-phosphate dehydrogenase (G6PD) deficiency. The goals of this study were (i) to evaluate the performance of a two-tier galactosemia screening protocol, (ii) to explore the impact on and acceptability to families of reporting G6PD deficiency as a secondary finding, and (iii) assess the communication and follow-up process for positive G6PD deficiency screening results. The two-tiered galactosemia approach increased the positive predictive value (PPV) for galactosemia from 8% to 79%. An additional 119 positive newborn screen results were reported for G6PD deficiency with a PPV of 92%. The results show that there may be utility in reporting G6PD deficiency results. Most parents who participated in the study reported having some residual worry around the unexpected diagnosis; however, all thought it was helpful to know of their child's diagnosis of G6PD deficiency. Finally, the communication process for reporting G6PD deficiency newborn screen results was determined to result in appropriate follow up of infants.

Effect of neonatal reticulocytosis on glucose 6-phosphate dehydrogenase (G6PD) activity and G6PD deficiency detection: a cross-sectional study

BackgroundScreening for G6PD deficiency in newborns can help prevent severe hemolysis, hyperbilirubinemia, and bilirubin encephalopathy, as recommended by the World Health Organization (WHO). It has been speculated that the presence of a high number of reticulocytes in newborns interferes with the diagnosis of G6PD deficiency since reticulocytes contain higher amounts of G6PD enzyme than mature erythrocytes. Therefore, the purposes of this study were to assess the effect of reticulocytosis in the determination of blood G6PD activity in Thai newborns by using a novel automated UV-based enzymatic assay and to validate the performance of this assay for the detection of G6PD deficiency in newborn samples.MethodsThe levels of reticulocytes and G6PD activity were measured in blood samples collected from 1,015 newborns. G6PD mutations were identified using TaqMan® SNP genotyping assay, PCR–restriction fragment length polymorphism (PCR–RFLP), and direct sequencing. The correlation between the levels of reticulocytes and G6PD activity was examined. The performance of the automated method was compared with that of the fluorescent spot test (FST) and the standard quantitative assay.ResultsThe automated assay detected G6PD deficiency in 6.5% of the total newborn subjects compared to 5.3% and 6.1% by the FST and the standard method, respectively. The minor allele frequencies (MAFs) of G6PD ViangchanG871A, G6PD MahidolG487A, and G6PD UnionC1360T were 0.066, 0.005, and 0.005, respectively. The reticulocyte counts in newborns with G6PD deficiency were significantly higher than those in normal male newborns (p < 0.001). Compared with normal newborns after controlling for thalassemias and hemoglobinopathies, G6PD-deficient patients with the G6PD ViangchanG871A mutation exhibited elevated reticulocyte counts (5.82 ± 1.73%, p < 0.001). In a group of G6PD normal newborns, the percentage of reticulocytes was positively correlated with G6PD activity (r = 0.327, p < 0.001). However, there was no correlation between G6PD activity and the levels of reticulocytes in subjects with G6PD deficiency (r = -0.019, p = 0.881). The level of agreement in the detection of G6PD deficiency was 0.999, while the area under the receiver operating characteristic (AUC) curve demonstrated that the automated method had 98.4% sensitivity, 99.5% specificity, 92.4% positive predictive value (PPV), 99.9% negative predictive value (NPV), and 99.4% accuracy.ConclusionsWe report that reticulocytosis does not have a statistically significant effect on the detection of G6PD deficiency in newborns by both qualitative and quantitative methods.

Hemolytic anemia in a 2-years old caused by Glucose 6 Phosphate Dehydrogenase Deficiency (G6PD): a case report

Background: Glucose-6-Phosphate-Dehydrogenase (G6PD) deficiency is an enzyme abnormality related to sex chromosomes (x-linked), which is inherited. The activity or stability of the G6PD enzyme decreases, thus causing the breakdown of red blood cells when an individual is exposed to exogenous substances that can cause oxidative damage. This case study aims to describe a rare case of hemolytic anemia caused by glucose 6 phosphate dehydrogenase deficiency in a two-year-old boy. Case Presentation: A two-year-old boy presented with agitated, pale, yellowish eyes and body and projectile vomiting after consuming fava beans. He had a prior history of packed red cell transfusion. Physical examination found the conjunctivae were pale with the icteric sclera. Laboratory examination revealed leukocytosis, moderate normochromic normocytic anemia, increased reticulocytes count, and elevated total and unconjugated bilirubin. Peripheral blood smear examination found microcytes, spherocytes, fragmentocytes, and burr cells. Coomb test was negative with decreased G6PD result. Conclusion: G6PD deficient patients have no symptoms until exposed to oxidizing drugs, infections or after fava beans ingestion which may cause hemolytic anemia and jaundice. Several methods of screening and diagnosis of G6PD can be performed to avoid morbidity and mortality.

Deficiência da glicose-6-fosfato desidrogenase (G6PD) em recém-nascidos, numa maternidade pública da cidade de Campina Grande, Estado da Paraíba, Brasil

Nos eritrócitos deficientes de glicose 6 fosfato desidrogenase (G6PD), a diminuição da redução de NADP em NADPH, leva a um baixo potencial redutor que interfere na capacidade metabólica oxidativa do organismo, ficando vulnerável a hemólise, com oxidação da membrana do glóbulo, podendo levar a crise hemolíticas. As manifestações da deficiência de G6PD são desencadeadas por fatores ambientais como drogas, fava, infecções. As manifestações clínicas consistem de crise hemolítica e icterícia neonatal. Estudos indicam que no Brasil a prevalência de G6PD está em torno de 1 a 10%, sendo os maiores índices em populações com ancestralidade africana. O objetivo foi a pesquisa da deficiência da G6PD em recém-nascidos numa maternidade pública na cidade de Campina Grande. Trata-se de um estudo transversal, onde foram coletados sangue do cordão umbilical de 147 recém-nascidos. O diagnóstico da deficiência da G6PD será realizado através do teste da redução da metahemoglobina (teste de Brewer). A cidade de Campina Grande, bem como todo o Compartimento da Borborema não apresentam nenhum dado relativo a prevalência da deficiência da G6PD. O Ministério da Saúde fortalece através do SUS a coleta de informações relativas a esta doença na perspectiva de implantação de um programa de atenção à saúde dos doentes detectados, promovendo melhoras na sua qualidade de vida.

Frequency of Human Paraoxonase-1 Q192R Polymorphism and Measurement of Oxidative Stress Parameters in Infants with G6PD Deficiency

Background and Aims: This study aimed to investigate the frequency of Q192R polymorphism and oxidative stress markers in infants with glucose-6phosphate dehydrogenase (G6PD) deficiency.&#x0D; Materials and Methods: This is a case-control study in which 60 male infants (2-4 months old) with G6PD deficiency along with 60 age- and sexmatched healthy neonates were included. The diagnosis of G6PD deficiency was made by Beutler test by which the G6PD enzyme activity is measured by the fluorescent spot test. The blood samples were taken from all infants, and the sera were isolated for the evaluation of Paraoxonase-1 (PON1) and malondialdehyde (MDA) using the spectrophotometric method. Restriction fragment length polymorphism was applied for determination of Q192R polymorphism (rs 662).&#x0D; Results: The frequencies of QQ, QR, and RR genotypes were 55%, 39%, and 6%, respectively in infants with G6PD deficiency while the above genotype frequencies were 45%, 49%, and 6%, respectively in healthy neonates. The frequency of R and T alleles failed to show any significant difference when G6PD deficient infants and healthy neonates were compared. The results indicated PON1 activity and MDA levels being significantly (p&lt;0.05) higher in neonates with G6PD deficiency compared with their healthy counterparts.&#x0D; Conclusion: Contrary to previous studies, it was indicated that the presence of RQ and RR genotypes at Q192R position is associated with decreased activity of PON1 and increased oxidative stress. In this study, no significant differences were found in the genotype and allele frequency of PON1 Q192R polymorphism between the case and control groups. Also, this frequency was not consistent with the results obtained from oxidative stress conditions.

Fine Mapping of Glucose 6 Phosphate Dehydrogenase (G6PD) Deficiency in a Rural Malaria Area of South West Odisha Using the Clinical, Hematological and Molecular Approach.

IntroductionThe aim of the study was to enumerate the clinical, hematological, and molecular spectrum of G6PD deficiency in malaria endemic regions of south west Odisha.MethodsDiagnosis of G6PD deficiency was made by using the Di-chloroindophenol Dye test in two south west districts (Kalahandi and Rayagada) of Odisha State. Demographic and clinical history was taken from each individual using a pre-structured questionnaire. Molecular characterization of G6PD deficiency was done using PCR-RFLP and Sanger sequencing.ResultsA total of 1981 individuals were screened; among them, 59 (2.97%) individuals were G6PD deficient. The analysis revealed that G6PD deficiency was more among males (4.0%) as compared to females (2.3%). Prevalence of G6PD deficiency was significantly higher among tribal populations (4.8%) as compared to non-tribal populations (2.4%) (p=0.012, OR=2.014, 95%CI=1.206–3.365). Twenty four individuals with G6PD deficiency had mild to moderate anemia, whereas 26 G6PD deficient individuals had a history of malaria infection. Among them, 3 (11.5%) required blood transfusion during treatment. Molecular analysis revealed G6PD Orissa as the most common (88%) mutation in the studied cohort. G6PD Kaiping (n=3), G6PD Coimbra (n=2) and G6PD Union (n=1) were also noted in this cohort.ConclusionThe cumulative prevalence of G6PD deficiency in the present study is below the estimated national prevalence. G6PD deficiency was higher among tribes as compared to non-tribes. Clinical significance for G6PD deficiency was noted only in malaria infected individuals. Rare G6PD Kaiping and G6PD Union variants were also present.

Screening of Glucose-6-Phosphate Dehydrogenase Deficiency in Neonatal Hyperbilirubinaemia at 300-Bedded Pyin Oo Lwin General Hospital

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is one of the most common human enzyme deficiencies in the world. It is particularly common in populations living in malaria-endemic areas, affecting more than 400 million people worldwide. This hospital- and laboratory-based, cross-sectional descriptive study was conducted with the aim of determining the prevalence of G6PD deficiency among 200 newborns at 300-bedded Pyin Oo Lwin General Hospital during January to March 2017. The participants were 103 girls (58.5%) and 97 boys (41.5%). Both qualitative and quantitative measurements by using Brewer's method and G-SIX kit method were applied for diagnosis of G6PD deficiency. Total serum bilirubin level was measured by Bilirubinometer. Of the 200 newborns, 21(10.5%) were G6PD deficient. The overall prevalence of G6PD deficiency was 10.5% (21/200) and male was predominant than female (17.5% vs 3.9%). Out of 10.5% (21/100)G6PD deficient newborns, 5(23.8%) and 16(76.2%) were mild and moderate G6PD deficiency, respectively. Regarding hyperbilirubinaemia, 9(42.9%), 3(14.3%), 2(19.0%) and 5(23.8%) were severe, moderate and mild hyperbilirubinaemia and normal bilirubin, respectively. This study showed that a significant correlation between the severity of hyperbili- rubinaemia and G6PD activity (p &lt;0.05). Taking into consideration of the above results, the high prevalence can be useful for providing appropriate prevention and early treatment of complications in routine neonatal screening in this area.

The impact of using primaquine without prior G6PD testing: a case series describing the obstacles to the medical management of haemolysis

Radical cure of Plasmodium vivax malaria in glucose-6-phosphate dehydrogenase (G6PD) deficient individuals employs weekly primaquine dosing. This is the only recommended regimen for this patient sub-group. If national malaria programs mandate daily primaquine dosing (the recommended regimen for G6PD normal individuals), then G6PD testing before prescription is necessary to avoid iatrogenic haemolysis in G6PD deficient individuals. In this case series, two P. vivax infected patients with unknown G6PD status from two different countries were prescribed primaquine as per national malaria program guidelines. During treatment both patients presented to the clinic with symptoms of anaemia after taking primaquine incorrectly. The clinical management of the iatrogenic severe haemolysis that occurred in these patients demonstrates the various adverse effects primaquine can cause, that other common medical treatments also have haemolytic potential, and how the diagnosis of G6PD deficiency can be elusive during acute haemolysis. Health care providers should provide careful instructions about primaquine dosing, be watchful for haemolysis, and have a high index of suspicion for G6PD deficiency in the presence of haemolysis if the G6PD status is previously unknown.

Proteomics‐Based Investigations of Neglected and Tropical Diseases

Proteomics‐Based Investigations of Neglected and Tropical Diseases

Review and drug therapy implications of glucose-6-phosphate dehydrogenase deficiency.

The pathophysiology, diagnosis, and medication-use implications of glucose-6-phosphate dehydrogenase (G6PD) deficiency, the most common enzyme deficiency in humans, are reviewed. Originally identified as favism in patients who experienced hemolysis after ingestion of fava beans, G6PD deficiency results from an X-linked chromosomal mutation that leads to reduced activity of the enzyme responsible for the final step of the pentose phosphate pathway, through which reduced nicotinamide adenine dinucleotide phosphate required for protection of cells from oxidative stress is produced. G6PD deficiency affects about 400 million people worldwide. Diagnosis of G6PD can be made through detection of enzymatic activity (by spectrophotometric testing, fluorescence testing, or formazan-based spot testing) or molecular analysis to detect known mutations of the gene encoding G6PD. Most individuals with G6PD deficiency are asymptomatic throughout life. Symptoms of acute hemolysis associated with G6PD deficiency include anemia, fatigue, back or abdominal pain, jaundice, and hemoglobinuria. The most common precipitators of oxidative stress and hemolysis in G6PD deficiency include medication use and infection. G6PD deficiency should be considered in patients who experience acute hemolysis after exposure to known oxidative medications, infection, or ingestion of fava beans. A diagnosis of G6PD deficiency is most often made through enzymatic activity detection, but molecular analysis may be required in females heterozygous for the disorder. When clinically feasible, rasburicase, primaquine, dapsone, pegloticase, and methylene blue should not be used until a G6PD diagnostic test has been performed.

A study of glucose-6-phosphate dehydrogenase deficiency in neonatal hyperbilirubinemia

Background: Glucose-6-phosphate dehydrogenase (G6PD) is an enzyme essential for basic cellular functions, including protection of red cell proteins from oxidative damage. G6PD deficiency is the most common red cell enzyme abnormality associated with hemolysis as well as with neonatal jaundice, kernicterus, and even death. It plays a protective role against malaria. It is a genetically inherited sex-linked abnormality. Objectives: To determine the age, sex, and different community-wise incidence of G6PD deficiency in neonatal subjects having neonatal hyperbilirubinemia. Materials and Methods: This study was carried out among 170 neonates at Biochemistry Department of B.J. Medical College & Civil Hospital, Ahmedabad, Gujarat, India, between August 2007 and July 2009. Results: Of 170 neonates, 123 were males and 47 were females, and 18 neonates showed deficiency of G6PD in red cells. Parsi community had highest incidence of G6PD enzyme deficiency. Muslim community had higher incidence than Hindu community because of consanguineous marriages. Of total 18 G6PD enzyme–deficient subjects, 11 were early neonates and 7 were late neonates. Of these 11 G6PD enzyme–deficient early neonates, 5 were 6-day old. Highest incident was found on 2-day-old neonates. Conclusion: The G6PD-deficient neonates are prone to a greater incidence of neonatal jaundice. Methylene blue reduction test is useful screening procedure, so the quantitative enzyme assay must be used for a definitive diagnosis of G6PD deficiency.

Glucose-6-Phosphate Dehydrogenase Activity Levels in White Newborn Infants

To define normal levels of glucose-6-phosphate dehydrogenase (G6PD) activity in a population of North American white newborns. We studied 2 white newborn populations, ≥35weeks of gestation. In the retrospective study, G6PD activity was measured (on clinical indication) in 242 newborns aged ≤7days. In the prospective study, we measured G6PD activity in umbilical cord blood samples in 347 newborns and daily transcutaneous bilirubin levels in these infants. The mean G6PD activity level was 12.3±3.1 units per gram hemoglobin (U/gHb) in the retrospective population and 13.3±1.8 U/gHb in the prospective population, and there was no difference between males and females. The distribution of values suggested that infants with activity levels <7 U/gHb should be considered deficient and 8 infants (6 males and 2 females), all in the retrospective population had such levels. As in other ethnic populations, the mean G6PD activity in white newborn infants is substantially greater than that of white adults. The lower limits of normal are also similar to those of other newborn ethnic groups and of adults. The diagnosis of G6PD deficiency should be considered in any white infant whose G6PD activity is <7 U/gHb.

A novel cytofluorometric assay for the detection and quantification of glucose-6-phosphate dehydrogenase deficiency

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is an X-linked enzymopathy that affects hundreds of millions of people worldwide, conferring increased risk of neonatal jaundice and oxidant-induced hemolytic anemia. Screening and diagnosis of G6PD deficiency is currently performed using genetic or biochemical assays, the former being cost ineffective in populations with significant allelic heterogeneity, and the latter being limited in ability to detect female heterozygotes. Cytochemical assays can obviate these shortcomings, but at the expense of added technical complexity and labor. We describe here a simple, novel cytofluorometric method that extends the classic methemoglobin reduction test, assessing G6PD deficiency at the level of an individual erythrocyte. In preliminary testing in Malian children, there was strong concordance between our method and established genetic and biochemical techniques. The assay is robust and economical, and could serve as a screening method as well as a research tool, especially for high-throughput applications such as flow cytometry.

Aregenerative anemia and erythrocytes hemighosts: a case report

An 11 year old African boy without previous history was hospitalised for fever and a severe anaemia (haemoglobin = 55 g/L) with low reticulocyte count. Blood smear showed more than 35% of ghost red blood cells which allows the diagnosis of G6PD deficiency (< 1% of normal level). Anaemia was demonstrated as haemolytic and was associated with a drepanocytosis trait. Aspect of red blood cells on blood smear remains important for the diagnosis of congenital or acquired red blood cell diseases, even abnormalities are often of low specificity.

The effect of different haemoglobins on the glucose-6-phosphate dehydrogenase deficiency screening test (Brewer's test)

Glucose-6-phosphate dehydrogenase (G6PD) deficiency and haemoglobinopathies are common causes of haemolytic anaemia in Sri Lanka. Even though G6PD enzyme assay is indicated for diagnosis of G6PD deficiency, due to lack of reagents many of the government hospitals in Sri Lanka make the diagnosis only on the basis of a screening test (Brewer's test) for this deficiency. Both these tests are indicated in patients with chronic haemolytic anaemia. The aim of the study was to observe the effect of different haemoglobins on screening test for G6PD deficiency. We investigated the patients and carriers for commoner haemoglobinopathies (Hb E, Hb S, and Hb D) and thalassaemias using G6PD deficiency screening test as well as G6PD enzyme assay. The results show that patients with Hb E/β thalassaemia, Hb E trait and Hb E disease gave a false positive reaction with G6PD screening test (n=47), even though their G6PD enzyme levels were normal. Two subjects with Hb E showed low enzyme levels with positive screening test. Hb S (n=5) and Hb D (n=2) did not have any effect on the screening test for G6PD deficiency. The patients with Hb E and normal enzyme levels are likely to be misdiagnosed as having G6PD deficiency, if only the screening test is utilized. This study emphasizes the necessity for employing an enzyme assay method for the diagnosis of G6PD deficiency, at least in districts with high Hb E prevalence. Keywords: G6PD; Brewer's test; thalassaemia; haemoglobinopathy. DOI: 10.4038/jdp.v5i1.2957 Journal of Diagnostic Pathology 2006/07; 5: 30-34

New onset diabetes complicated by haemolysis and rhabdomyolysis: a case report and review of the literature

IntroductionPreviously undiagnosed glucose-6-phosphate dehydrogenase (G6PD) deficiency can be unmasked by a diabetic crisis and both can be associated with rhabdomyolysis. The relationship between diabetes and G6PD deficiency is discussed and the possible triggers for haemolysis as outlined in this case report. The incidence of G6PD deficiency is 10% in African-American males and up to 35% in parts of Africa so an increased awareness of G6PD deficiency is important when treating diabetes in these populations.Case presentationA 54-year-old Kenyan man presented with a 3-day history of reduced appetite, weakness and reduced level of consciousness as a result of a hyperglycaemic diabetic crisis with both hyperosmolarity and ketoacidosis. The patient then developed haemolysis and a raised creatine kinase level. A diagnosis of G6PD deficiency and rhabdomyolysis was made.ConclusionThis case highlights the importance of simple laboratory investigations in the early identification of the rarer complications of diabetic crisis such as haemolysis secondary to G6PD deficiency and rhabdomyolysis.

Biochemical and cytochemical evaluation of heterozygote individuals with glucose-6-phosphate dehydrogenase deficiency

The aim of this study was to diagnose heterozygous glucose-6-phosphate dehydrogenase (G6PD) deficient females by an inexpensive cytochemical G6PD staining method that is easy to perform, allowing diagnosis of G6PD deficiency without cumbersome genetic analysis. Three subject groups were included in the study. The first group consisted of 15 hemizygous deficient males. The second and the third group were composed of 15 heterozygous deficient females and 15 healthy individuals, respectively. Biochemical determination and cytochemical staining of G6PD activity were performed in samples of all subjects. Results obtained with the cytochemical staining method correlated significantly with the biochemical data (p < 0.001), but a only 51-68% of the erythrocytes were stained positively in females with normal biochemical G6PD activity despite their having a G6PD-deficient child. This observation clearly indicates that these individuals are heterozygously deficient. These findings show that the cytochemical staining method to detect G6PD activity in erythrocytes is reliable, sensitive and specific and is superior to the biochemical method. Therefore, this method can be used routinely to detect heterozygous G6PD deficiency.

Glucose-6-phosphate dehydrogenase activity in male premature and term neonates

Background: Glucose-6-phosphate dehydrogenase (G6PD) activity is higher in term neonates than in adults. Some studies have suggested that activity may be even higher in preterm infants. Objectives: To determine if...

Index of Suspicion

Index of Suspicion