Evaluation of Glucose-6-Phosphate Dehydrogenase Stability in Blood Samples under Different Collection and Storage Conditions

Castro, Simone Martins de; Weber, Raquel; Dadalt, Vivian; Santos, Vanessa F. C.; Reclos, George J.; Pass, Kenneth A.; Giugliani, Roberto

doi:10.1373/clinchem.2005.048520

Cited by 19 publications

(11 citation statements)

References 3 publications

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“…These are typically in low prevalence in the population and require significant screening to identify patients with these G6PD activity characteristics. Unfortunately, when these specimens are obtained they must be tested within a short time frame (seven to 14 days) [17]. Establishing a specimen bank enriched for specimens with intermediate G6PD activity, with deficient G6PD activity, and from confirmed heterozygous females would be of great value to both the development of tests for G6PD deficiency and to their early evaluation.…”

Section: Discussionmentioning

confidence: 99%

Cryopreservation of glucose-6-phosphate dehydrogenase activity inside red blood cells: developing a specimen repository in support of development and evaluation of glucose-6-phosphate dehydrogenase deficiency tests

Kahn

LaRue

Bansil

et al. 2013

Malar J

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BackgroundGlucose-6-phosphate dehydrogenase (G6PD) deficiency is a common human enzyme deficiency. It is characterized by abnormally low levels of G6PD activity. Individuals with G6PD deficiency are at risk of undergoing acute haemolysis when exposed to 8‒aminoquinoline-based drugs, such as primaquine. For this reason it is imperative to identify individuals with G6PD deficiency prior to administering these anti-malarial drugs. There is a need for the development and evaluation of point-of-care G6PD deficiency screening tests suitable for areas of the developing world where malarial treatments are frequently administered. The development and evaluation of new G6PD tests will be greatly assisted with the availability of specimen repositories.MethodsCryopreservation of erythrocytes was evaluated as a means to preserve G6PD activity. Blood specimens from 31 patients including ten specimens with normal G6PD activity, three with intermediate activity, and 18 with deficient activity were cryopreserved for up to six months.ResultsGood correlation in G6PD activity between fresh and cryopreserved specimens (R2 = 0.95). The cryopreserved specimens show an overall small drop in mean G6PD activity of 0.23 U/g Hb (P=0.23). Cytochemical staining showed that intracellular G6PD activity distribution within the red blood cell populations is preserved during cryopreservation. Furthermore, the mosaic composition of red blood cells in heterozygous women is also preserved for six months or more. The fluorescent spot and the BinaxNOW qualitative tests for G6PD deficiency also showed high concordance in G6PD status determination between cryopreserved specimens and fresh specimens.ConclusionsA methodology for establishing a specimen panel for evaluation of G6PD tests is described. The approach is similar to that used in several malaria research facilities for the cryopreservation of parasites in clinical specimens and axenic cultures. Specimens stored in this manner will aid both the development and evaluation of current and emerging G6PD tests. The availability of G6PD tests is a critical bottleneck to broader access to drugs that confer radical cure of Plasmodium vivax, a requirement for elimination of malaria.

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Section: Discussionmentioning

confidence: 99%

Cryopreservation of glucose-6-phosphate dehydrogenase activity inside red blood cells: developing a specimen repository in support of development and evaluation of glucose-6-phosphate dehydrogenase deficiency tests

Kahn

LaRue

Bansil

et al. 2013

Malar J

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“…Moreover, exposure to heat en route to a central laboratory may affect the accuracy of biochemical G6PD testing. 47 Whom to screen? A second challenge is whether screening should be (i) targeted to high-risk groups or (ii) universal.…”

Section: Research Gapsmentioning

confidence: 99%

Should we screen newborns for glucose-6-phosphate dehydrogenase deficiency in the United States?

et al. 2013

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Glucose-6-phosphate dehydrogenase (G6PD) deficiency, a common X-linked enzymopathy can lead to severe hyperbilirubinemia, acute bilirubin encephalopathy and kernicterus in the United States. Neonatal testing for G6PD deficiency is not yet routine and the American Academy of Pediatrics recommends testing only in jaundiced newborns who are receiving phototherapy whose family history, ethnicity, or geographic origin suggest risk for the condition, or for infants whose response to phototherapy is poor. Screening tests for G6PD deficiency are available, are suitable for use in newborns and have been used in birth hospitals. However, US birth hospitals experience is limited and no national consensus has emerged regarding the need for newborn G6PD testing, its effectiveness or the best approach. Our review of current state of G6PD deficiency screening highlights research gaps and informs specific operational challenges to implement universal newborn G6PD testing concurrent to bilirubin screening in the United States.

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“…Loss in enzyme activity had previously been found to be quite significant in blood samples (albeit anticoagulated with EDTA) stored at room temperature, when after 15-day storage, G6PD levels fell by approximately 60% [16]. While storage-dependent changes in G6PD function have been described in leucoreduced donor blood units [17] additives such as that described by Kahn et al, [18] appeared to extend the functionality of intact red cells when analysed by flow cytometry, even though the overall G6PD level remained stable over a 21-day period.…”

Section: Discussionmentioning

confidence: 96%

Abrogation of red blood cell G6PD enzyme activity through Heat treatment: development of survey material for the UK NEQAS G6PD scheme

Roper

Soni²,

Fletcher

et al. 2017

Int J Lab Hematology

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Summary Introduction Participation in external quality assessment (EQA) is central to the maintenance of high‐quality laboratory results in patient diagnosis and clinical trials. Laboratories in the TAF112582 DETECTIVE study (ClinicalTrials.gov identifier: NCT01376167) are enrolled in the United Kingdom National Quality Assessment Scheme (UK NEQAS) for glucose‐6‐phosphate dehydrogenase (G6PD) quantitative assay, which utilizes ovine (sheep) blood as a readily available source of apparently G6PD‐deficient survey material. A substitute for sheep blood was sought because some non‐UK sites in the study encountered participation difficulties due to the strict regulations on the import of sheep blood into their countries. Methods G6PD activity in normal human donor blood was abrogated by the action of heat under controlled conditions. Residual G6PD activity in the heated samples was measured by UK NEQAS using the Trinity Biotech 345 kit (Trinity Biotech) and a Jenway 6715 UV/Vis spectrophotometer with external temperature control to monitor enzyme kinetics and linearity over a set time. Heat‐treated material was also assayed for G6PD activity and assessed for its acceptability as EQA survey material by selected UK laboratories. Results Blood heated at 45 °C for 15 h showed a reduction in G6PD activity of 76.3 ± 4.6% (n = 6) and was considered acceptable as EQA material in terms of appearance and behaviour by the majority of UK sites in the trial. Conclusions We have developed a simple heat‐treatment procedure to produce EQA survey material with low/intermediate G6PD activity, similar to that found in females heterozygous for G6PD deficiency.

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Evaluation of Glucose-6-Phosphate Dehydrogenase Stability in Blood Samples under Different Collection and Storage Conditions

Cited by 19 publications

References 3 publications

Cryopreservation of glucose-6-phosphate dehydrogenase activity inside red blood cells: developing a specimen repository in support of development and evaluation of glucose-6-phosphate dehydrogenase deficiency tests

Cryopreservation of glucose-6-phosphate dehydrogenase activity inside red blood cells: developing a specimen repository in support of development and evaluation of glucose-6-phosphate dehydrogenase deficiency tests

Should we screen newborns for glucose-6-phosphate dehydrogenase deficiency in the United States?

Abrogation of red blood cell G6PD enzyme activity through Heat treatment: development of survey material for the UK NEQAS G6PD scheme

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