Delay in the measurement of eosin-5′-maleimide (EMA) binding does not affect the test result for the diagnosis of hereditary spherocytosis

Ciepiela, Olga; Kotuła, Iwona; Górska, Elżbieta; Stelmaszczyk-Emmel, Anna; Popko, Katarzyna; Szmydki‐Baran, Anna; Adamowicz-Salach, Anna; Demkow, Urszula

doi:10.1515/cclm-2012-0240

Cited by 11 publications

(9 citation statements)

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“…EMA binding test is used as a screening test to diagnose HS [ 3 , 7 – 16 ]. The result of the test is calculated as a decrease in the fluorescence of EMA-bound red blood cells of patients compared to EMA fluorescence of six healthy reference samples.…”

Section: Discussionmentioning

confidence: 99%

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Mean corpuscular volume of control red blood cells determines the interpretation of eosin-5′-maleimide (EMA) test result in infants aged less than 6 months

et al. 2015

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Eosin-5′-maleimide (EMA) binding test is a flow cytometric test used to detect hereditary spherocytosis (HS). To perform the test sample from patients, 5–6 reference samples of red blood are needed. Our aim was to investigate how the mean corpuscular volume (MCV) of red blood cells influences on the value of fluorescence of bounded EMA dye and how the choice of reference samples affects the test result. EMA test was performed in peripheral blood from 404 individuals, including 31 children suffering from HS. Mean fluorescence channel of EMA-RBCs was measured with Cytomics FC500 flow cytometer. Mean corpuscular volume of RBCs was assessed with LH750 Beckman Coulter. Statistical analysis was performed using Graph Pad Prism. The correlation Spearman coefficient between mean channel of fluorescence of EMA-RBCs and MCV was r = 0.39, p < 0.0001. Interpretation of EMA test depends on MCV of the reference samples. If reference blood samples have lower MCV than the patients MCV, EMA test result might be negative. Due to different MCV values of RBCs in infancy and ca. Three months later, EMA test in neonates might be interpreted falsely negative. Samples from children younger than 3 months old had EMA test result 86.1 ± 11.7 %, whereas same samples that analyzed 4.1 ± 2.1 later had results of 75.4 ± 4.5 %, p < 0.05. Mean fluorescence of EMA-bound RBC depends on RBC’s volume. MCV of reference samples affects EMA test results; thus, we recommend selection of reference samples with MCV in range of ±2 fL compared to MCV of patient RBC’s.

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Section: Discussionmentioning

confidence: 99%

“…The procedure of the test was according to that described previously [ 16 ]. Five microliters of whole blood from each sample was washed with 0.9 % NaCl solution and incubated in darkness at room temperature for 1 h with 25 μL of EMA dye (0.5 mg/mL, in phosphate-buffered saline, PBS; Fluka, Gillingham, UK), with intermittent mixing.…”

Section: Methodsmentioning

confidence: 99%

Mean corpuscular volume of control red blood cells determines the interpretation of eosin-5′-maleimide (EMA) test result in infants aged less than 6 months

et al. 2015

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“…Immediate hypersensitivity can be explored in functional assays demonstrating basophil degranulation . MFC can also provide useful information on red blood cells in cases of suspected spherocytosis or to detect the presence of fetal erythrocytes . Finally, MFC enables for some platelet disorders to be explored .…”

Section: Introductionmentioning

confidence: 99%

Flow cytometry in hematological nonmalignant disorders

Béné

Bris

Robillard

et al. 2015

Int J Lab Hematology

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Summary Multiparameter flow cytometry (MFC) has become an integral part of the diagnosis and classification of hematological malignancies. However, several nonmalignant or premalignant disorders may benefit from this technology in hematology laboratories. This review provides information on the normal immunophenotypic characteristics of peripheral blood leukocyte subsets and their modifications in several clinical conditions. The usefulness of MFC and the specific markers that can be investigated in hyperlymphocytosis, infection, hypereosinophilia, paroxysmal nocturnal hemoglobinuria, and large granular lymphocyte disorders is described. Mention is also made of the developments of MFC for analyses of red blood cells or platelets.

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“…If the diagnosis is unclear, confirmatory testing may be done using the eosin-5'-maleimide (EMA) binding test, which is highly sensitive and specific for hereditary spherocytosis 15. This is a quick and reliable flow cytometric test that uses EMA as a form of dye that binds to plasma membrane proteins, allowing for easy recognition of membrane defects 14. Osmotic fragility testing also can be used to confirm the diagnosis, but lacks the sensitivity and specificity of the EMA binding test and can be particularly inaccurate in neonates 14…”

Section: Discussionmentioning

confidence: 99%

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2023

Jaapa

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Delay in the measurement of eosin-5′-maleimide (EMA) binding does not affect the test result for the diagnosis of hereditary spherocytosis

Abstract: Results of the EMA binding test do not depend on storage time of stained samples when stored at 4°C up to 24 h after staining.

Cited by 11 publications

References 25 publications

Mean corpuscular volume of control red blood cells determines the interpretation of eosin-5′-maleimide (EMA) test result in infants aged less than 6 months

Mean corpuscular volume of control red blood cells determines the interpretation of eosin-5′-maleimide (EMA) test result in infants aged less than 6 months

Flow cytometry in hematological nonmalignant disorders

What is making this newborn excessively sleepy?

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