Glucose-6-phosphate Dehydrogenase Deficiency Research Articles

Malaria remains one of the most significant parasitic diseases globally, exerting a disproportionate burden in sub-Saharan Africa, Asia, and Latin America. Host genetic factors play a critical role in determining susceptibility to malaria infection and severity, with evolutionary evidence showing that Plasmodium parasites have shaped human genetic diversity over centuries. Protective adaptations such as hemoglobinopathies, G6PD deficiency, and variations in erythrocyte surface antigens demonstrate the selective pressure malaria has exerted on human populations. Advances in genomics and molecular biology have expanded understanding of single-nucleotide polymorphisms (SNPs), copy number variations (CNVs), and immune-related polymorphisms that modulate host-parasite interactions. This paper reviews major genetic determinants of malaria susceptibility, explores their implications for pathophysiology, and highlights the role of modern genomic tools in uncovering novel protective and risk alleles. By integrating genetic insights with public health strategies, research in this field offers significant opportunities for vaccine development, targeted therapies, and precision medicine approaches in the global fight against malaria. Keywords: Malaria susceptibility, Host genetics, Hemoglobinopathies, Single nucleotide polymorphisms (SNPs) and Copy number variations (CNVs).

Malaria remains one of the world’s most devastating infectious diseases, caused by Plasmodium parasites and transmitted through Anopheles mosquitoes. Despite repeated exposure in endemic regions, the development of long-lasting immunity remains elusive due to complex host–parasite interactions, immune evasion mechanisms, and antigenic variation. This paper examines the multifaceted immunological responses to malaria, emphasizing how Plasmodium falciparum manipulates host immunity through antigenic variation, immune modulation, and persistence mechanisms that prevent sterilizing immunity. The roles of humoral and cellular immune responses, particularly memory B cells, T follicular helper (Tfh) cells, and tissue-resident memory T cells (Trm), are discussed in relation to their contribution to long-term protection. Global initiatives such as the Roll Back Malaria campaign, WHO’s Global Technical Strategy for Malaria (2016–2030), and community-based engagement efforts highlight the progress and challenges in malaria control and eradication. Advances in vaccine research, including whole-organism vaccines, recombinant platforms, and bispecific antibody therapies are reshaping prospects for immunological protection. However, ethical considerations concerning human challenge trials, gene-drive technology, and the management of G6PD deficiency remain critical in global malaria research. Understanding the molecular mechanisms underlying immune memory, immune evasion, and vaccine-induced immunity is pivotal to accelerating the development of next-generation vaccines and innovative therapeutics essential for malaria elimination and eventual eradication. Keywords: Malaria Immunology, Antigenic Variation, Immune Memory, Vaccine Development and Global Eradication Efforts.

Schizophrenia is a severe mental disorder characterized by aberrant brain development, influenced by genetic and environmental factors, with an incompletely understood etiology. Glucose-6-phosphate dehydrogenase (G6PD), a critical enzyme in the pentose phosphate pathway (PPP), facilitates NADPH production for biosynthesis and redox homeostasis. Recent studies suggests that PPP inhibition and consequent oxidative stress contribute to schizophrenia pathogenesis. While clinical investigations have proposed a link between G6PD deficiency and schizophrenia, the underlying biological mechanisms remain unestablished. We here demonstrate that brain-specific G6PD knockout induces schizophrenia-like behaviors in mice, establishing a direct association between G6PD deficiency and schizophrenia. Proteomic analysis revealed aberrant synaptic protein expression in the knockout mice. These mice also exhibited synaptic impairments, including reduced presynaptic vesicles and diminished dendritic spines. Our findings suggest that G6PD deficiency disrupts synaptic homeostasis, contributing to schizophrenia-like behaviors. Our study provides novel insights into the molecular mechanisms of schizophrenia, identifying G6PD as an important regulator of synaptic function and a potential therapeutic target.

Imported malaria is a threat to malaria elimination. A case investigation (CI) with reactive active case detection (RACD) strategy is commonly used for the detection of malaria infections around the index cases. CI with RACD was conducted for detection of subpatent malaria among febrile mobile and migratory population (MMP) individuals around index cases in the state of Punjab in India, by microscopy, rapid diagnostic tests (RDTs) and nested PCR (nPCR). Glucose-6-phosphate dehydrogenase deficiency (G6PDd) was determined in Plasmodium vivax-positive patients. A total of 119 malaria cases were reported, out of which 93.3% were P. vivax (111/119) and 6.7% (8/119) were Plasmodium falciparum. CI with RACD was initiated around 28 MMP index cases positive by RDT and microscopy. A total of 176 fingerprick samples were collected from febrile MMP individuals, from which 11 (11/176; 6.3%) infected individuals were detected: eight by RDT, microscopy and nPCR, and three (3/11; 27.3%) subpatent cases by only nPCR. None of the positive patients were found positive for G6PDd. Subpatent malaria in MMP individuals can pose a challenge in malaria elimination. The RACD strategy around index cases may help to reduce the risk of spillover in the local population.

This study aimed to explore reference interval (RI) for the determination of glucose-6-phosphate dehydrogenase (G6PD) deficiency in adults by means of G6PD/6PGD ratio assay. Given that G6PD/6PGD exhibits differences between male and female, the currently available RI of G6PD/6PGD is not applicable for screening G6PD deficiency in local population. This study investigates RI for the G6PD/6PGD in Yangjiang, providing valuable insights for clinical decision-making and treatment.A total of 5,186 individuals were included in this study, comprising 2,386 males and 2,800 females. G6PD/6PGD of these individuals was determined. G6PD genotypes and thalassemia genotypes of different groups were diagnosed, followed by relevant statistical analyses, and RI was determined using a non-parametric approach, defined by the 2.5th and 97.5th percentiles of the distribution.In Yangjiang, RI for G6PD/6PGD was 1.32-2.08 in male and 1.19-2.03 in female. The adjusted male median (AMM) and the normal male median (NMM) values were 1.68 and 1.69, respectively. Among the population with G6PD gene mutations, the upper limit of G6PD/6PGD RI was 0.5 for male. A 45% (G6PD/6PGD = 0.7605) threshold of the NMM could be used to estimate the probability of G6PD variants. When G6PD/6PGD was 0.7605, male had a variant-positivity rate of 97.39% and female was 92.31%.The G6PD/6PGD ratio measurements in this study enable more accurate assessment of G6PD activity levels in the Yangjiang population. These findings provide clinically relevant insights for optimizing treatment strategies and enhancing the prediction and management of G6PD deficiency.

Disclosure: B. Agrawal: None. S. nagpaul: None. S. Prabhakaran: None. S.A. Narra: None. H. Patel: None.Introduction: Diabetic ketoacidosis (DKA), a life-threatening complication more common in type 1 diabetes, marked by hyperglycemia, ketosis, and anion gap metabolic acidosis due to insulin deficiency. This triggers fat breakdown and ketone production. Such metabolic derangements can precipitate or exacerbate hemolysis, potentially revealing underlying conditions like G6PD deficiency. Case Presentation: A 25-year-old male with newly diagnosed insulin-dependent diabetes mellitus presented with two days of nausea, vomiting, abdominal and chest pain, polyuria, and polydipsia, following insulin non-compliance. He was tachycardic, tachypneic, and hypothermic on admission. Labs showed severe DKA (glucose 620 mg/dL, bicarbonate 5, anion gap 34, beta-hydroxybutyrate 14), HbA1c >15%, hyponatremia (125), hyperkalemia (5.9), leukocytosis (WBC 27), and elevated lactate (3.5), suggestive of pneumonia. Urinalysis revealed proteinuria and glycosuria. He was admitted to the ICU and treated with IV fluids, bicarbonate, insulin drip, and antibiotics. During hospitalization, he developed normocytic anemia with hemoglobin dropping from 13 to 6.8 g/dL, requiring transfusion. Hemolysis workup revealed elevated bilirubin, LDH, reticulocyte count, decreased haptoglobin, schistocytes on peripheral smear, and a normal hemoglobin electrophoresis. G6PD assay confirmed G6PD deficiency. After resolution of DKA, he was transitioned to subcutaneous insulin and discharged with folic acid, insulin therapy, and education on avoiding G6PD triggers. He was advised hematology and endocrinology follow-up. Discussion: G6PD deficiency compromises red blood cell (RBC) defense against oxidative stress due to impaired NADPH and glutathione production. DKA-induced acidosis and hyperglycemia increase oxidative stress, promoting RBC fragility and hemolysis. Dehydration and hyperosmolarity further reduce RBC survival. Hemolysis can present with nonspecific symptoms overlapping with DKA, delaying diagnosis. Laboratory findings include anemia, jaundice, elevated indirect bilirubin, LDH, reticulocytes, and low haptoglobin. A significant hemoglobin drop during DKA warrants investigation. A negative Coombs test helps rule out autoimmune causes, while schistocytes suggest oxidative injury. G6PD levels may appear normal during acute hemolysis due to reticulocytosis; retesting may be needed after recovery. Management includes supportive care, transfusions if needed, and avoiding oxidative triggers. Patient education and strict insulin adherence are vital to prevent recurrence. Conclusion: This case illustrates how DKA-induced oxidative stress can unmask G6PD deficiency, leading to hemolysis. Clinicians should evaluate for hemolytic anemia in DKA with unexplained anemia, especially in high-risk populations, and consider inherited enzymopathies as part of the differential.Presentation: Sunday, July 13, 2025

G6PD deficiency classically presents with hemolytic anemia, but associations have been described with atherogenesis and coronary artery disease secondary to increased oxidative stress. There are limited data that also suggest a link between G6PD deficiency and the development of congenital heart disease (CHD), possibly through the same mechanism. We hypothesize a higher incidence of CHD among individuals with G6PD deficiency as well as worse hospital outcomes for those with CHD and G6PD deficiency. We performed a retrospective review of a national administrative database from 10/2019 to 3/2024 for admissions of individuals aged 0-25 years with ICD-10 codes for moderate-to-severe CHD with and without G6PD deficiency (D55.0, D75.A). Additional data included demographics, hospital outcomes, and costs. Hospital outcomes were compared. There were 213,708 admissions with CHD and <1% had G6PD deficiency. Of the 6,363 admissions with G6PD, 289 (4.5%) had CHD and of 11,179,670 without G6PD, 213,419 (1.9%) had CHD (p < 0.001). Admissions with CHD and G6PD deficiency were predominantly Black/African American and male (consistent with G6PD deficiency epidemiology), with a longer length of stay and higher hospital costs than those without G6PD deficiency. Our findings support a higher incidence of CHD associated with G6PD deficiency as well as worse hospital outcomes for those with CHD and comorbid G6PD deficiency. More work is needed to identify a potential causative link to further understand mechanisms for CHD development.

Background:Glucose-6-phosphate dehydrogenase deficiency (G6PDd) is a common genetic disorder that can lead to life-threatening complications, including acute hemolytic anemia. Effective screening using advanced technology, such as point-of-care test (POCT), is critical to ensure proper clinical management, particularly in malaria-endemic areas. However, economic evidence of the quantitative POCT, such as cost structure, cost-effectiveness, and potential areas for cost savings, particularly in developing countries, including Malaysia, is scarce.Methods:A top–down and time-driven activity-based costing was conducted at the Canselor Tuanku Muhriz Hospital, Malaysia, between February 1 and 28, 2023, for the CareStart™ Biosensor S1 (CBS1) and compared with the OSMMR-2000D test, focusing on the duration to perform the screening test for G6PDd (i.e. cost per minute of turnaround time). The activities, items, and individuals involved were identified using the activity-based costing form, which includes a workflow and detailed breakdown tables. The capital cost, recurrent cost, and total expenditure for both methods were determined. Univariate sensitivity analysis was conducted by adjusting several key variables, such as human resources, disposable items, and equipment costs, by a 20% increase or decrease, and analyzing their effect on the overall cost structure.Results:In 2023, the cost of G6PDd screening using the CBS1 in Malaysia was RM47.54 (USD11.15) per test, compared with a higher cost of RM246.27 (USD54.18) per test with the OSMMR-2000D. It has been demonstrated that the major cost division for CBS1 was in the disposable item cost (58.18%), which includes the device’s strips and reagents, whereas the major cost division for OSMMR-2000D was in the transportation (59.72%). The univariate sensitivity analysis revealed that the disposable item cost for CBS1 was the most sensitive parameter, which affected the total expenditure the most.Conclusion:This study provides valuable economic evidence on the efficiency of quantitative POCT for G6PDd. In countries with high G6PD prevalence as in Malaysia, such screening efficiency is crucial, as it would not only improve patient outcomes but also significantly reduce overall healthcare expenditures. This analysis proved that G6PDd screening using POCT is more cost-effective and time-efficient, and the method should be a policy priority for rapid, mobile, and remote screening of targeted populations.

Innate immune response to bacterial infections in hospitalized glucose-6-phosphate dehydrogenase-deficient patients; diagnostic value of white blood cell count.

BackgroundAdding single low-dose (0.25 mg/kg) primaquine (SLDPQ) to block Plasmodium falciparum transmission is now a WHO recommendation. Whether SLDPQ increases haemolysis in glucose-6-phosphate dehydrogenase deficient (G6PDd) patients, leading to increased folate demand and impaired haemoglobin (Hb) recovery is unknown. This study sought to answer this question.MethodsThis randomized, placebo-controlled trial measured serial plasma folate concentrations [Day (D) 0, 3, 7 and 28] in falciparum-infected Ugandan and Congolese children (6 months to 11 years), treated with age-dosed SLDPQ/placebo and artemether-lumefantrine/dihydroartemisinin-piperaquine. Genotyping defined G6PD (G6PD c.202T allele) status. Multiple linear and non-linear, mixed effects, cubic spline regression were fitted to identify factors significantly associated with plasma folate at baseline and over time, respectively.Results408 children (3 had missing D0 values) had ≥ 1 plasma folate value. Of these, 66 (16.2%) were G6PD-deficient, 51 (12.5%) heterozygous females, 283 normal and 8 unknown. Mean baseline folate concentrations were 10.83 [standard deviation (SD) 3.58, SLDPQ] vs 10.92 (SD 4.54, placebo) ng/ml, associated independently with baseline Hb [estimate: 0.52 ng/ml (95% CI: 0.26 to 0.79, p = 0.0001)] and baseline parasitaemia [estimate: − 0.18 ng/ml (− 0.32 to − 0.05, p = 0.007)]. For all patients, mean plasma folate concentration paralleled mean haemoglobin concentration with an initial mean fall of 1.65 ng/ml (p < 0.0001 vs. baseline), followed by a sustained rise achieving a mean D28 concentration of 11.04 (SD 4.45) ng/ml. Over time, only age (p = 0.0001), male sex (p = 0.017) and baseline parasitaemia (p = 0.029) were significantly associated with a reduced plasma folate.ConclusionSLDPQ and G6PD status did not compromise posttreatment plasma folate concentrations in young children with acute uncomplicated falciparum malaria, providing additional evidence of SLDPQ safety and supporting its use without G6PD testing.Trial registration The trial is registered, reference number ISRCTN11594437.

BackgroundOutside of sub-Saharan Africa, Plasmodium vivax has become the dominant species of malaria. Focal mass drug administration (fMDA) is a potential strategy to support elimination efforts, but controlled studies are lacking.MethodsThe FocaL mass drug Administration for Plasmodium vivax Malaria Elimination (FLAME) study is a 3-year cluster randomized controlled trial to determine the impact and safety of fMDA to reduce P. vivax transmission. The study will be conducted in Loreto, Peru, where standard interventions have reduced P. vivax cases, but transmission persists due to a high proportion of subclinical infections. Thirty low transmission communities (API < 250 cases/1000 population) will be randomized 1:1 to fMDA versus control using a restricted randomization. All communities will receive Peruvian national standard malaria control measures. In the intervention arm, high-risk individuals (living within 200 m of a P. vivax case reported in the prior 2 years) without contraindication to study medications, including G6PD deficiency, will receive three cycles of fMDA over a 2-year period. Each cycle will include two rounds of directly observed therapy delivered 2 months apart. The fMDA regimen will include 25 mg/kg chloroquine (CQ) plus a single 300 mg dose of tafenoquine (TQ) for individuals age ≥ 16 years, and 25 mg/kg of CQ plus 7 days of 0.5 mg/kg/day of primaquine (PQ) if younger. The primary outcome is the cumulative incidence of symptomatic P. vivax malaria. The sample size provides 80% power to detect at least a 68% relative reduction in cumulative P. vivax incidence, based on alpha of 0.05 and a coefficient of variation (k) of 0.87. Secondary outcomes include safety, cost-effectiveness, and infection prevalence and seroprevalence which will be assessed in annual cross-sectional surveys. Safety will be assessed in passive and active pharmacovigilance, including post-treatment screening for G6PD-associated hemolysis by assessing for anemia and hematuria in a sample.DiscussionThe trial will generate evidence regarding fMDA for P. vivax and inform malaria elimination efforts in Peru and similarly endemic settings. Findings will be disseminated in peer-reviewed publications and through stakeholder meetings in Peruvian and international research forums.Trial registrationClinicaltrials.gov NCT05690841. This trial was registered on 09 January 2023. Peruvian Clinical Trial Registry (REPEC) 020–23. This trial was registered on 21 February 2024.Supplementary InformationThe online version contains supplementary material available at 10.1186/s13063-025-09112-1.

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a genetic condition caused primarily by single nucleotide polymorphisms (SNPs). It is the most prevalent enzymopathy globally and directly impacts the ability of red blood cells to respond to oxidative stress. This study aimed to determine the prevalence of G6PD variants in Amazon blood donors. A cross-sectional study was conducted at the Foundation of Hemotherapy and Hematology of Amazonas (HEMOAM) between January and June of 2024. The study analyzed 5,000 donor samples using the quantitative polymerase chain reaction (qPCR) to detect G6PD genetic variants. G6PD variants were present in 364 (7.28%) donors, with the most common being c.376A>G (3.82%), c.292G>A (1.92%), and simultaneous c.292G>A/376A>G (0.90%). Other rare variants were also identified, including c.653C>T, c.1093G>A, c.968T>C; c.632A>T; c.1429G>A; c.700G>T; c.1192G>A and c.197T>A/c.202G>A, representing 0.64% of the total. Most donors exhibiting mutations were male (71.7%), with a mean age of 34.1±11.9 years for males and 34.0±10.5 years for females. The most prevalent blood group in both sexes was O+, with a frequency of 59.7% in males and 61.9% in females. This study constitutes the most extensive molecular testing for G6PD mutations in the state of Amazonas, offering a comprehensive overview of G6PD deficiency (G6PDd) in the region. The estimated prevalence rate of G6PD mutations among Manaus blood donors was 7.28%, which is consistent with other findings in Brazil. In conclusion, G6PDd has the potential to compromise red blood cell quality during storage and their survival post-transfusion. This study enhanced the methods used to identify mutations impacting blood storage, with the aim of improving transfusion safety and efficacy.

Glucose-6-phosphate dehydrogenase deficiency (G6PDd) is one of the most common genetic disorders worldwide and remains highly prevalent in malaria-endemic regions. Individuals with G6PDd are at risk of severe complications, including acute haemolytic anaemia, when exposed to oxidative triggers. In Malaysia, the Proto-Malay Orang Asli (PMOA), the second largest indigenous group in Peninsular Malaysia, represents a vulnerable population. This study aimed to estimate the prevalence and mutation spectrum of G6PDd in this community. A total of 258 peripheral blood samples (91 males, 167 females) were screened using a quantitative G6PD assay (OSMMR2000-D). DNA from 73 samples was genotyped with the Hybribio G6PD GenoArray test, and 39 underwent targeted sequencing. The adjusted male median (AMM) of G6PD activity was 9.6 U/gHb (95% CI: 8.9–10.3 U/gHb), with 30% and 80% thresholds corresponding to 2.9 and 7.7 U/gHb, respectively. At the 30% cut-off threshold, the overall estimated prevalence of G6PDd was 6.8% (16/237; 12 males and 4 females). A total of 21 subjects were G6PD-intermediate (7 males and 14 females), and the remaining 221 subjects were G6PD-normal (72 males and 150 females). Genotyping identified 18 hemizygous males, 13 heterozygous females, and 3 homozygous females. Five G6PD variants were detected: G6PD Viangchan (39.5%), G6PD Coimbra (28.9%), G6PD Union (23.7%), G6PD Kaiping (5.3%), and rs782038151 (2.6%). This study demonstrates that G6PDd is common in the PMOA population, with notable molecular diversity. These findings have important implications for malaria control and the safe use of antimalarial drugs in this high-risk community.

Glucose-6-phosphate dehydrogenase deficiency (G6PDd) is one of the most common genetic disorders worldwide and remains highly prevalent in malaria-endemic regions. Individuals with G6PDd are at risk of severe complications, including acute haemolytic anaemia, when exposed to oxidative triggers. In Malaysia, the Proto-Malay Orang Asli (PMOA), the second largest indigenous group in Peninsular Malaysia, represents a vulnerable population. This study aimed to estimate the prevalence and mutation spectrum of G6PDd in this community. A total of 258 peripheral blood samples (91 males, 167 females) were screened using a quantitative G6PD assay (OSMMR2000-D). DNA from 73 samples was genotyped with the Hybribio G6PD GenoArray test, and 39 underwent targeted sequencing. The adjusted male median (AMM) of G6PD activity was 9.6 U/gHb (95% CI: 8.9-10.3 U/gHb), with 30% and 80% thresholds corresponding to 2.9 and 7.7 U/gHb, respectively. At the 30% cut-off threshold, the overall estimated prevalence of G6PDd was 6.8% (16/237; 12 males and 4 females). A total of 21 subjects were G6PD-intermediate (7 males and 14 females), and the remaining 221 subjects were G6PD-normal (72 males and 150 females). Genotyping identified 18 hemizygous males, 13 heterozygous females, and 3 homozygous females. Five G6PD variants were detected: G6PD Viangchan (39.5%), G6PD Coimbra (28.9%), G6PD Union (23.7%), G6PD Kaiping (5.3%), and rs782038151 (2.6%). This study demonstrates that G6PDd is common in the PMOA population, with notable molecular diversity. These findings have important implications for malaria control and the safe use of antimalarial drugs in this high-risk community.

Medicinal plants, as an effective source of traditional medicine, have genuine utility, and about 80% of the suburban population relies on plants as primary health care globally, especially in Africa, due to their availability and fewer complications. This study was designed to determine the antimicrobial effect of Azadirachta indica extracts in the context of G6PD deficiency and malaria co-infection on clinical isolates, comprising five (5) bacteria and four (4) fungi, using the agar well diffusion method. Minimum inhibitory and bactericidal concentration, including kinetic growth of the isolates, were determined by macrodilution and spectrophotometry methods. The aim of the study is to determine antimicrobial activities and phytochemical constituents of Azadirachta indica (Neem) on multidrug-resistant clinical isolates. The phytochemical and functional group profiling in the extracts were performed using GC–MS standard method and Fourier-Transform Infrared (FTIR) spectrophotometry. In-silico molecular docking analysis of the bioactive compounds in neem extract and oil was determined by computational modelling tools and the molecular Auto Dock software. Data were analysed using one-way ANOVA to compare the mean levels of significance of the parameters, where the level of significance was set as (P&lt;0.05), and Duncan Multiple Range Test (DMRT) was used to compare the significance between the groups. The results showed that aqueous neem demonstrates narrow antimicrobial potential, while methanol, including oil of neem extracts, displayed broad antimicrobial potential, and aqueous neem gave the strongest inhibitory zone of 14 mm against Escherichia coli. Neem extracts demonstrate broad antifungal potential over amphotericin B, while methanolic neem has a 0.60 mm wide inhibitory zone against Saccharomyces cerevisiae. The minimum inhibitory concentration and minimum bactericidal concentration for neem oil were 2.00 mg/ml stronger. Growth kinetics indicated bacteriostatic effects of neem extracts on Staphylococcus aureus, Escherichia coli, Candida albicans, and Aspergillus niger. Neem oil and powder contain 14 and 18 phytochemical profiles while α-D-Glucopyranose and n-Hexadecanoic acid were identified at the highest peak area of 21.98 % and 12.31% respectively. In-silico molecular docking identified diethylphthalate as a strong microbial protein inhibitor at -8.5 kcal, low binding energy with S aureus 1txt and E. coli 2ZIP proteins, while 9,17-octadecadienal (Z), methyl 10-methyl hexadecanoate, and oleic acid contributed to neem oil’s antibacterial activity with low binding energy of -8.00 kcal with S aureus 1txt and E. coli 2ZIP proteins. However, the therapeutic importance of neem in combating pathogenic microbes and their sustainability as an alternative medicine is significant. The findings of this research acknowledge and scientifically validate the use of plants against microbial pathogenic activities.

The Impact of G6PD Deficiency on Liver Damage and Disease Progression in Hepatocellular Carcinoma: A Cross-Sectional Study in Thailand

Abstract Background Hemoglobin A1c (HbA1c) is a widely used lab test to assist in diagnosing and managing diabetes. However, in patients with known disorders of red blood cell turnover and certain hemoglobin variants, this value is inaccurate. Current American Diabetes Association guidelines state that HbA1c should not be used to diagnose diabetes in these individuals. A 2021 publication by Sivasanskar showed there is a significant quality gap with inappropriate HbA1c test orders among those with sickle cell disease (SCD) at hundreds of healthcare facilities across the United States. We suspect this is also the case for glucose 6-phosphate dehydrogenase (G6PD) deficiency and thalassemias known to affect red blood cell turnover. Multiple recent publications have shown that HbA1c levels at all levels of glucose are much lower for those with G6PD deficiency than for controls. Since both SCD and G6PD deficiency are known to occur in African-Americans to a greater extent than Caucasians, we believe this could be a significant issue in our health care system leading to delayed diagnosis or inappropriate management of diabetes in this population. Methods Using retrospective electronic health record data from one children’s hospital, one additional hospital and several outpatient clinics in our health system, we examined &amp;gt;40,000 orders for glucose and alternate markers from 1955 unique patients with an ICD-10 code for SCD, thalassemia, or anemia caused by enzyme deficiency. The protocol for the study was approved by the Washington University IRB. The majority of these patients were diagnosed with sickle-cell disease or trait (79%) while only 11 (&amp;lt;1%) were diagnosed with G6PD deficiency and the remainder (20%) with thalassemia. Results Of the total orders examined, 95% were for glucose (either alone or as part of a panel with additional tests). For the remaining 5% of orders, 447 were for glucose tolerance testing (gestational), 21 for glucose tolerance testing (non-gestational), 1487 for HbA1c, and 55 for fructosamine. Twenty-nine percent of patients with SCD or trait had at least one HbA1c order and this percentage was higher (51%) in patients with a thalassemia. For the 11 patients with G6PD deficiency, 36% had a HbA1c order. Fructosamine orders occurred for only 25 unique patients, none of whom had G6PD or other anemia of enzyme disorders. Less than 1% patients with thalassemia and only 1.4% of patients with SCD or trait had an order for fructosamine. For the patients in whom fructosamine was ordered, 80% also had one or more HbA1c orders, including nine instances with both tests ordered during the same week. Conclusion The percentage of patients with a disorder of red blood cells and HbA1c orders is significant while fructosamine is rarely ordered. This suggests opportunities for electronic ordering interventions and/or additional provider education regarding appropriate use of alternate markers of glucose control in this group of patients.

Abstract Background Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common inherited enzyme deficiency. It is a risk factor for kernicterus (i.e., bilirubin induced neurological damage) in jaundiced newborns and hemolysis following oxidant stress exposure. In June 2022, New York State (NYS) mandated hospitals to provide newborn screening for G6PD deficiency by a quantitative enzyme method or gene sequencing for newborns with certain clinical presentations, however since most hospital laboratories do not offer quantitative enzyme testing, samples are sent to reference laboratories. Reference laboratory turnaround time (TAT) is not optimal for making urgent clinical decisions for treating jaundice and providing results prior to newborn discharge. The Baebies FINDER G6PD Test (FINDER G6PD) provides G6PD enzyme activity values within 17 minutes using 50 µL of whole blood, providing timely results for all patients. Methods Validation of FINDER G6PD included intra-run precision studies with 20 runs of 2 levels of quality control (QC), and establishing a reference range by testing 120 healthy individuals. TAT, the time from sample collection to result reporting, and the prevalence of G6PD deficiency in the patient population were determined for two 6-month intervals: December 1, 2023 – May 31, 2024 when samples were sent to the reference laboratory and July 1, 2024 – December 31, 2024, when samples were tested in the hospital laboratory with FINDER G6PD. Results Intra-run precision performed with normal level QC demonstrated a mean of 12.7 U/gHb, standard deviation (SD) of 0.47, and a coefficient of variation (CV) of 3.7%. Intra-run precision performed with the abnormal (low) QC demonstrated a mean of 5.6 U/gHb, SD of 0.18, and CV of 3.3%. A total of 120 individuals, 62 females, and 58 males, were recruited for the reference range study, however, 2 samples were excluded because they demonstrated deficiency. Based on the remaining 118 samples, the reference interval was determined to be 8.0 U/gHb – 15.2 U/gHb using the transformed parametric method in EP Evaluator. The median TAT for reference laboratory testing was 75.4 hours (3.1 days) compared to 4.5 hours (0.2 days) for FINDER G6PD in-hospital laboratory testing. The prevalence of G6PD deficiency, defined as a test result below the reference range, was 15.5% (43/277) for all patients tested and 11.1% (2/18) for newborns (i.e., patients &amp;lt;=1 month) in reference laboratory testing and 17.9% (43/240) for all patients tested and 14.3% (3/21) for newborns in FINDER G6PD in-hospital laboratory testing. Conclusion The prevalence of G6PD deficiency in the population served by New York Presbyterian Hospital-Columbia University Irving Medical Center is high, demonstrating the need for G6PD testing results for patient care. Hospital laboratory testing using FINDER G6PD provides results in 4.5 hours compared to 3.1 days ensuring G6PD diagnosis prior to hospital discharge of newborns. This rapid TAT enables real-time decision-making allowing timely interventions, including treatment and education for G6PD-deficient newborns. Other patients, such as those being evaluated for possible treatment with medications contraindicated in individuals with G6PD deficiency (e.g., rasburicase), also benefit from more rapid testing results.

Abstract Background Glucose-6-phosphate Dehydrogenase (G6PD) testing is used for diagnosis of G6PD deficiency. World Health Organization (WHO) advises that an individual’s G6PD status should be determined to guide dose and duration of anti-malarial drugs, and there are still unmet needs for G6PD testing to early screen neonates. The existing G6PD screening methods usually require multiple steps of manual operation. The results of these methods are often impacted by the variability or errors in manual operation. Therefore, they are difficult to accurately identify the population with G6PD mutation. This study evaluated the performance of a novel fully-automated G6PD/Hb detection method (G6PDH), and compared its diagnostic value with other methods, so as to provide a more precise technical means for the early screening and diagnosis of the disease. Methods In this study, the automatic lysis mode of Mindray BS-2800M analyzer was used to realize the G6PD/Hb twin-tests in parallel, in which G6PD and Hb (hemoglobin) were detected simultaneously using the same blood sample, and the G6PD/Hb ratio was calculated, defined as G6PDH. The age and gender specific reference intervals with this method were established. Based on the results of Multiplex Melting Curve Analysis (MMCA, a genetic mutation detecting method), the diagnostic efficacy and accordance rate of G6PDH and other biochemical methods such as G6PD/6PGD, G6PD single enzyme method were analyzed. Results According to the guidance of WHO, the reference intervals of G6PDH in non-neonates were established as &amp;lt;2.31, =2.31, 2.31∼6.17, =6.17 for deficient males/females, normal males, intermediate females, and normal females. For neonates, they were &amp;lt;4.14, =4.14, 4.14∼11.03, =11.03, respectively. Compared with the results of MMCA method on 185 samples, the sensitivity and accuracy of the G6PDH method were 93.5% and 91.7% respectively, which were significantly higher than those of G6PD/6PGD method (74.0%, 74.4%) and G6PD single enzyme method (68.3%, 67.8%). The Youden index of G6PDH method was 0.81, higher than that of 0.74 and 0.63 by other two methods, respectively. And the missed diagnosis rate was 6.5%, lower than that of the other two methods (26.0%, 31.7%). Moreover, in 64 female patients with heterozygous mutation, the diagnose accordance rate of G6PDH was 87.5%, while the other two was 51.6% and 39.1%. Conclusion Mindray provides an innovative automated G6PDH method on BS-2800M to accurately identify G6PD deficiency population, and significantly reduces the missed diagnosis rate of high-risk population with G6PD deficiency, especially for female heterozygous mutation. The G6PDH method could be well used for the screening, diagnosis and monitoring of G6PD deficiency, and could be considered as the preferred method in clinical laboratory.

Abstract Background G6PD deficiency is an X-linked, hereditary genetic defect caused by mutations in the G6PD gene. It is a common human enzyme defect and is prevalent in some regions of the world. The most common clinical manifestations of G6PD deficiency are neonatal jaundice and acute hemolytic anemia that is often triggered by an exogenous agent. Currently, the mainstream methods for detecting G6PD deficiency are the fluorescent-spot-method and the UV-enzymatic method running on biochemistry analyzers. The fluorescent-spot-method is not able to identify heterozygotes G6PD deficiency, with long test time and low efficiency. Traditional UV-enzymatic method detects G6PD only, requiring pre-manual sample-dilution and treatment to obtain hemolysate. These operations are cumbersome and time-consuming. G6PD mostly resides in red cells. Due to the variation of the red cell quantity in samples, the G6PD deficiency needs to be reported as the ratio of G6PD activity over Hb (U/g). Commonly, the Hb concentration from the same sample is measured separately by a hematology analyzer. The ratio G6PD/Hb (U/g) is then calculated, which is not convenient if there is a large demand for G6PD tests. This study evaluated the performances of a new G6PD assay with enhanced reagent formulation for stability on Mindray BS-600M analyzer having on-board sample hemolysis and twin-chemistries (G6PD and Hb) measured in parallel. The ratio of G6PD/Hb is reported directly by the analyzer. Methods Mindray G6PD assay is a UV-enzymatic method. It is designed with an innovative approach with on-board hemolysis on the BS-600M system, using one method to measure G6PD activity and a 2nd method to measure Hb in parallel to report two results simultaneously and calculated the ratio of G6PD/Hb (U/g). Within 15 min, the ratio results can be obtained, which greatly improves the work efficiency compared with the 1-3h required by the traditional methods. The reagent onboard stability, using two levels of quality controls and serum pools, was conducted with a constant reagent consumption mode to simulate as it is used in clinical labs. Following CLSI protocols, the assay performances were evaluated: method comparison, onboard stability, precision, and linearity. Results On Mindray BS-600M analyzer, our G6PD(U/g) assay correlated well with the results using traditional UV-enzymatic method reagent with manual sample pre-treatment for obtaining hemolysate (Mindray = 1.0191* Manual method-0.0934, r=0.95). Mindray G6PD assay also demonstrated an excellent onboard stability. With uncapped reagent on the system, the reagent stability is for 21 days without recalibration (with less than ± 5% deviation from day zero results). In comparison, a routinely used UV-enzymatic reagent (freeze-dried) from another company showed significant drifting (deviation &amp;gt;±10% vs that on day 0, within 7 days). The repeatability and with-lab CVs of our assay were ranged from 0.40% to 1.86%, respectively. Conclusion On Mindray BS-600M analyzer, the new G6PD assay shown characteristics of the short test time, high efficiency, excellent precision, and good correlation (accuracy). Therefore, we conclude that the new Mindray G6PD assay is suitable for use in routine clinical laboratories where large quantity of the G6PD tests are required.