Reliability of EMA Binding Test in the Diagnosis of Hereditary Spherocytosis in Italian Patients

D’Alcamo, Elena; Agrigento, Veronica; Sclafani, Serena; Vitrano, Angela; Cuccia, Liana; Maggio, Aurelio; Perrotta, Silverio; Capra, Marcello; Rigano, Paolo

doi:10.1159/000322253

Cited by 13 publications

(9 citation statements)

References 14 publications

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“…A correlation has been confirmed in individuals with high MCV due to overconsumption of alcohol, but in DAT-positive hemolytic anemia no correlation between MCV and MCF was observed [3]. Low MCV due to iron deficiency does not affect EMA MCF [3,6,20]. The results of the present study indicate no abnormality of the erythrocyte membrane in CDA III and show that standard flow cytometry cannot be used to discriminate between CDA III and normal controls.…”

Section: Discussionsupporting

confidence: 50%

Erythrocyte Flow Cytometric Analysis in Congenital Dyserythropoietic Anemia Type III-Evaluation of Eosin-5´-Maleimide, CD55, and CD59

Liljeholm¹

2013

J Blood Disord Transfus

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Section: Discussionsupporting

confidence: 50%

Erythrocyte Flow Cytometric Analysis in Congenital Dyserythropoietic Anemia Type III-Evaluation of Eosin-5´-Maleimide, CD55, and CD59

Liljeholm¹

2013

J Blood Disord Transfus

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“…4,5,16,17,19,37,38 Among the diagnostic methods considered, the recently proposed EMA-binding test is certainly the most interesting one, and it is being increasingly used by specialized laboratories because of its high sensitivity and specificity. 12,18,[23][24][25][26][27][28][29] This method directly targets the structural lesion of the disease, since the fluorescent probe eosin-5'-maleimide interacts with transmembrane proteins band 3, Rh protein, Rh glycoprotein and CD47 which are reduced in red cells from patients with hereditary spherocytosis; 30 defects of other cytoskeletal proteins, such as spectrin and protein 4.2, also induce a decrease in fluorescence intensity, likely because they create a long-range modulation effect on the dye binding site in band 3 protein. 39 The sensitivity of the EMA-binding test in this series is higher than that recently reported by Crisp et al, 12 and similar to 18 and Girodon et al 25 Interestingly, sensitivity is independent of clinical phenotype, being high also in patients with compensated anemia.…”

Section: Discussionmentioning

confidence: 99%

“…[13][14][15] More recently, the cryohemolysis test, 16,17 based on the observation that red cells from patients with hereditary spherocytosis are particularly susceptible to cooling at 0°C in hypertonic conditions, and the flow cytometric analysis of eosin-5'-maleimide-labeled intact red blood cells (EMA-binding test) 18 have been proposed as new methods for identifying hereditary spherocytosis. 19 The latter in particular has been proven to be a sensitive and specific diagnostic test for hereditary spherocytosis 12,18,[20][21][22][23][24][25][26][27][28][29] directly targeting the structural lesion of this disease, since the fluorescent probe, eosin-5'-maleimide, interacts with the protein band 3 complex. 30 The performance of the available direct or indirect diagnostic tests has been mostly evaluated individually and on limited number of cases.…”

Section: Introductionmentioning

confidence: 99%

Diagnostic power of laboratory tests for hereditary spherocytosis: a comparison study in 150 patients grouped according to molecular and clinical characteristics

Bianchi¹,

Fermo²,

Vercellati³

et al. 2011

Haematologica

152

140

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BackgroundThe laboratory diagnosis of hereditary spherocytosis commonly relies on NaCl-based or glycerol-based red cell osmotic fragility tests; more recently, an assay directly targeting the hereditary spherocytosis molecular defect (eosin-5'-maleimide-binding test) has been proposed. None of the available tests identifies all cases of hereditary spherocytosis. Design and MethodsWe compared the performances of the eosin-5'-maleimide-binding test, NaCl-osmotic fragility studies on fresh and incubated blood, the glycerol lysis test, the acidified glycerol lysis test, and the Pink test on a series of 150 patients with hereditary spherocytosis grouped according to clinical phenotype and the defective protein, with the final aim of finding the combination of tests associated with the highest diagnostic power, even in the mildest cases of hereditary spherocytosis. ResultsThe eosin-5'-maleimide-binding test had a sensitivity of 93% and a specificity of 98% for detecting hereditary spherocytosis: the sensitivity was independent of the type and amount of molecular defect and of the clinical phenotype. The acidified glycerol lysis test and Pink test showed comparable sensitivity (95% and 91%) . The sensitivity of NaCl osmotic fragility tests, commonly considered the gold standard for the diagnosis of hereditary spherocytosis, was 68% on fresh blood and 81% on incubated blood, and further decreased in compensated cases (53% and 64%, respectively). The combination of the eosin-5'-maleimide-binding test and acidified glycerol lysis test enabled all patients with hereditary spherocytosis to be identified. The eosin-5'-maleimide-binding test showed the greatest disease specificity. ConclusionsEach type of test fails to diagnose some cases of hereditary spherocytosis. The association of an eosin-5'-maleimide-binding test and an acidified glycerol lysis test enabled identification of all patients with hereditary spherocytosis in this series and, therefore, represents a currently effective diagnostic strategy for hereditary spherocytosis including mild/compensated cases.

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“…In laboratories with the ability to perform FACS-based studies, EMA binding has high sensitivity and specificity and is simple and rapidly performed. 29 EMA-binding results are not influenced by shipping or storage for several days, and may be suitable for study of patients who have been recently transfused.…”

Section: Hereditary Spherocytosismentioning

confidence: 99%

Abnormalities of the Erythrocyte Membrane

Gallagher

2013

Pediatric Clinics of North America

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Synopsis Primary abnormalities of the erythrocyte membrane, including the hereditary spherocytosis and hereditary elliptocytosis syndromes, are an important group of inherited hemolytic anemias. Classified by distinctive morphology on peripheral blood smear, these disorders are characterized by clinical, laboratory, and genetic heterogeneity. Among this group, hereditary spherocytosis patients are more likely to experience symptomatic anemia. Treatment of hereditary spherocytosis with splenectomy is curative in most patients. Once considered routine, growing recognition of the longterm risks of splenectomy, including cardiovascular disease, thrombotic disorders, and pulmonary hypertension, as well as the emergence of penicillin-resistant pneumococci, a concern for infection in overwhelming postsplenectomy infection, have led to re-evaluation of the role of splenectomy. Current management guidelines acknowledge these important considerations when entertaining splenectomy and recommend detailed discussion between health care providers, patient, and family. The hereditary elliptocytosis syndromes are the most common primary disorders of erythrocyte membrane proteins. However, most elliptocytosis patients are asymptomatic and do not require therapy.

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Reliability of EMA Binding Test in the Diagnosis of Hereditary Spherocytosis in Italian Patients

Cited by 13 publications

References 14 publications

Erythrocyte Flow Cytometric Analysis in Congenital Dyserythropoietic Anemia Type III-Evaluation of Eosin-5´-Maleimide, CD55, and CD59

Erythrocyte Flow Cytometric Analysis in Congenital Dyserythropoietic Anemia Type III-Evaluation of Eosin-5´-Maleimide, CD55, and CD59

Diagnostic power of laboratory tests for hereditary spherocytosis: a comparison study in 150 patients grouped according to molecular and clinical characteristics

Abnormalities of the Erythrocyte Membrane

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