Usefulness of the eosin‐5′‐maleimide cytometric method as a first‐line screening test for the diagnosis of hereditary spherocytosis: comparison with ektacytometry and protein electrophoresis

Girodon, François; Garçon, Loïc; Bergoin, E.; Largier, Marie; Delaunay, Jean; Fénéant-Thibault, Madeleine; Maynadié, Marc; Couillaud, Gérard; Moreira, Sophie; Cynober, Thérèse

doi:10.1111/j.1365-2141.2007.06944.x

Cited by 68 publications

(78 citation statements)

References 15 publications

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“…Thirty-two samples from non-related HS subjects were submitted to SDS-PAGE for red cell membrane protein analysis. The EMA binding test was performed as previously described and results expressed as proposed by Girodon et al 4,5 The distribution of the results obtained in the HS population compared to hemolytic diseases not related to red cell membrane disorder and controls is shown in Figure 1. Since different cut-off values have been used in previously published series (11% by Bianchi et al, 16% and 21% by Girodon et al) our results were stratified in 4 categories: under 11%, between 11 and 15%, between 16 and 20% and over 21% (Table 1).…”

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confidence: 99%

Testing for hereditary spherocytosis: a French experience

et al. 2012

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In a recent article, Bianchi et al. have compared several laboratory tests for diagnosis of hereditary spherocytosis (HS) by studying 150 HS patients.1 They concluded that the association of the eosin-5-maleimide (EMA) binding test and the acidified glycerol lysis test (AGLT) could identify all HS patients and represents an effective diagnostic tool. According to a receiver operating characteristic (ROC) curve analysis, the EMA cut-off value or the optimum fluorescence decrease that discriminates HS patients from controls was 11%. In these conditions, the test specificity was 98% and sensitivity 93%. In 93% of the cases, sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) identified the molecular defect.Osmotic gradient ektacytometry has demonstrated unequalled sensitivity for HS diagnosis, and easily discriminates between distinct red cell membrane disorders. It is not, however, readily available.2,3 We have used osmotic gradient ektacytometry to diagnose hemolytic diseases with suspected red cell membrane defects and compared these results with those of the EMA binding test. Eighty-two consecutive HS patients from 78 distinct families originating from different parts of France with typical ektacytometry curve and negative direct antiglobulin test were tested in parallel with 134 controls, 35 other red cell membrane disorders (7 congenital dyserythropoietic anemias type II (CDAII), 7 hereditary elliptocytosis, one pyropoikilocytosis, 4 Southeast Asian ovalocytosis (SAO), 16 xerocytosis) and 78 hemolytic syndromes with normal ektacytometric results. Thirty-two samples from non-related HS subjects were submitted to SDS-PAGE for red cell membrane protein analysis. The EMA binding test was performed as previously described and results expressed as proposed by Girodon et al. 4,5 The distribution of the results obtained in the HS population compared to hemolytic diseases not related to red cell membrane disorder and controls is shown in Figure 1. Since different cut-off values have been used in previously published series (11% by Bianchi et al., 16% and 21% by Girodon et al.) our results were stratified in 4 categories: under 11%, between 11 and 15%, between 16 and 20% and over 21% (Table 1).1,5 Overall, 85% of the HS cases showed a decrease in EMA binding of over 16% (with 72% over 21%). Only 4% were found in the 11-15% range and, notably, 11% of the HS displayed normal EMA binding. One control was in the 11-15% range. EMA binding test was normal in all 16 tested xerocytosis samples. As previously described, a significant decrease in EMA binding was also observed in other red cell membrane disorders, namely SAO, CDAII, elliptocytosis and pyropoikilocytosis. Careful examination of blood films can identify pyropoikilocytosis, elliptocytosis and SAO but not CDAII, frequently associated with spherocytes and that can be misdiagnosed for HS. Out of the 32 HS samples tested by SDS PAGE, the protein defect was identified in 23 (72%) cases, with 9 (39%) spectrin, 8 band 3 (35%), 5 (22%) ankyrin and one pr...

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Testing for hereditary spherocytosis: a French experience

et al. 2012

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“…Исследование сохранности цитоскелета эритроцитов методом проточной цитометрии -альтернативная мето-дика с высокой специфичностью и чувствительностью; ее эффективность в верификации наследственного сферо-цитоза подтверждена независимыми исследователями [7][8][9]12]. В нашем исследовании эффективность метода проточной цитометрии с применением эозин-5 малеими-да как диагностического теста для верификации диагноза наследственного сфероцитоза подтверждена для пациен-тов групп взрослого и детского возраста, за исключением пациента, перенесшего трансфузию за несколько дней до проведения исследования.…”

Section: Discussionunclassified

“…Незначительная часть красителя связывается с CD47, RH-ассоциированными белками. Снижение уров-ня экспрессии э5м на эритроцитах у пациентов с наслед-ственным сфероцитозом отражает дефицит мембранных белков, включая белок полосы 3, спектрин и белок поло-сы 4.2 [8,9].…”

Section: образцы кровиunclassified

Identification and Verification of Hereditary Spherocytosis by Means of Laboratory Diagnosis

Prokhorova¹,

Зуева²,

Соколова³

et al. 2014

Pediatr. farmakol.

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“…60 La prueba es indudablemente positiva para ESH si presenta una disminución de fluorescencia superior al 21%; sin embargo, el valor de corte a partir del cual la prueba es considerada negativa es variable, por lo que cada laboratorio debe establecer su propio valor. 8,56,60,74 Fragilidad osmótica eritrocitaria por citometría de flujo: Esta prueba cuantifica los eritrocitos resistentes a la hemólisis que se genera por el agregado de agua a una suspensión de sangre en solución fisiológica. Es decir que se evalúa el porcentaje de eritrocitos remanentes, en lugar de evaluar la hemólisis producida en el medio hipotónico.…”

Section: Prueba De Autohemólisisunclassified