{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,5]],"date-time":"2026-10-05T23:48:08Z","timestamp":1791244088986,"version":"4.1.0"},"reference-count":18,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2026,4,27]],"date-time":"2026-04-27T00:00:00Z","timestamp":1777248000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,4,27]],"date-time":"2026-04-27T00:00:00Z","timestamp":1777248000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["European J of Haematology"],"published-print":{"date-parts":[[2026,8]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>Hereditary spherocytosis (HS) is a congenital hemolytic anemia caused by erythrocyte membrane protein defects. While the eosin\u20105\u2032\u2010maleimide (EMA) binding test by flow cytometry is the established first\u2010line assay, this study evaluates its optimal cut\u2010offs and performance within a large, real\u2010world cohort.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>We retrospectively analyzed 181 consecutive samples (2017\u20132024) with suspected HS. EMA binding was performed by flow cytometry using conventional and standardized protocols; the latter (implemented in 2023) utilized fixed erythrocyte counts and FL1 bead calibration. Accuracy was assessed against a composite clinical\u2013laboratory reference standard. ROC analysis determined sensitivity, specificity, likelihood ratios, and AUC.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>HS was confirmed in 50\/181 cases (27.6%). EMA fluorescence reduction was significantly greater in HS (median 23.0%) than in non\u2010HS samples (9.0%). The optimal cut\u2010off (16.5%) yielded 96.0% sensitivity, 96.9% specificity, and AUC\u2009=\u20090.990 (95% CI 0.979\u20130.998). Both conventional and standardized protocols showed equivalent accuracy (AUC 0.988 vs. 0.992). False positives were mainly associated with severe microcytosis or autoimmune hemolytic anemia.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>In a consecutive, real\u2010world cohort of patients with suspected hereditary spherocytosis referred to a single reference laboratory receiving samples from across the country, the EMA test demonstrated high diagnostic accuracy consistent with previous reports. Importantly, implementation of a standardized protocol resolved reproducibility issues in fluorescence intensity measurement without compromising diagnostic performance, supporting its robustness and applicability in routine clinical laboratories.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1111\/ejh.70204","type":"journal-article","created":{"date-parts":[[2026,4,28]],"date-time":"2026-04-28T05:01:01Z","timestamp":1777352461000},"page":"459-466","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Evaluation of the Eosin\u20105\u2032\u2010Maleimide Flow Cytometry Test for the Diagnosis of Hereditary Spherocytosis"],"prefix":"10.1111","volume":"117","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-2776-9348","authenticated-orcid":false,"given":"Pablo Adrian","family":"Biaggioni","sequence":"first","affiliation":[{"name":"Hematology Department and Teaching Department IACA Laboratorios  Bah\u00eda Blanca Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alejandro Ceferino","family":"L\u00f3pez Romero","sequence":"additional","affiliation":[{"name":"Hematology Department IACA Laboratorios  Bah\u00eda Blanca Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,4,27]]},"reference":[{"key":"e_1_2_12_2_1","doi-asserted-by":"publisher","DOI":"10.3324\/haematol.2016.142463"},{"key":"e_1_2_12_3_1","doi-asserted-by":"publisher","DOI":"10.1111\/ijlh.12335"},{"key":"e_1_2_12_4_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365\u20102141.2011.08921.x"},{"key":"e_1_2_12_5_1","doi-asserted-by":"publisher","DOI":"10.7759\/cureus.70308"},{"key":"e_1_2_12_6_1","doi-asserted-by":"publisher","DOI":"10.1111\/ijlh.14376"},{"key":"e_1_2_12_7_1","doi-asserted-by":"publisher","DOI":"10.5045\/br.2013.48.4.237"},{"key":"e_1_2_12_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/jcla.24034"},{"key":"e_1_2_12_9_1","doi-asserted-by":"publisher","DOI":"10.1159\/000322253"},{"key":"e_1_2_12_10_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365\u20102141.2003.04730.x"},{"key":"e_1_2_12_11_1","first-page":"19","article-title":"El Test de 5\u2032\u2010Eosin\u2010Maleimide (EMA) en el Diagn\u00f3stico de la Esferocitosis Hereditaria: Revisi\u00f3n y Fundamentos","volume":"2","author":"Biaggioni P.","year":"2016","journal-title":"Informe ALAC"},{"issue":"3","key":"e_1_2_12_12_1","first-page":"307","article-title":"Esferocitosis Hereditaria: Experiencia Cl\u00ednica y Diagn\u00f3stica en Argentina","volume":"51","author":"Crisp R. L.","year":"2017","journal-title":"Acta Bioqu\u00edmica Cl\u00ednica Latinoamericana"},{"key":"e_1_2_12_13_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00277\u2010015\u20102377\u20100"},{"key":"e_1_2_12_14_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12288\u2010017\u20100907\u20108"},{"key":"e_1_2_12_15_1","volume-title":"R: A Language and Environment for Statistical Computing","author":"R Core Team","year":"2024"},{"key":"e_1_2_12_16_1","volume-title":"R Code for Diagnostic Accuracy Evaluation of the 5\u2032\u2010Eosin\u2010Maleimide (EMA) Flow Cytometry Test for Hereditary Spherocytosis\u2014IACA Study","author":"Biaggioni P. 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