{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,13]],"date-time":"2026-05-13T05:15:08Z","timestamp":1778649308405,"version":"3.51.4"},"reference-count":39,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,5,12]],"date-time":"2026-05-12T00:00:00Z","timestamp":1778544000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,5,12]],"date-time":"2026-05-12T00:00:00Z","timestamp":1778544000000},"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":["Cytometry Part B Clinical"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Hereditary spherocytosis (HS) is the most common congenital red blood cell membrane disorder, characterized by structural protein defects that lead to hemolytic anemia. Although several diagnostic tests exist, including osmotic fragility tests (OFTs), acidified glycerol lysis test (AGLT), and the EMA\u2010binding test (EMA), each presents specific limitations regarding sensitivity, specificity, or technical requirements. Flow cytometric osmotic fragility testing (OFT\u2010FCM) emerges as a promising complementary assay, offering a standardized workflow and rapid turnaround time. We conducted a retrospective study including 106 subjects (20 HS patients and 86 healthy controls) recruited at Hospital Cl\u00ednic de Barcelona between September 2024 and September 2025. Clinical and laboratory data were collected, and all participants underwent OFT, AGLT, EMA, and OFT\u2010FCM using two acquisition protocols (300 and 214\u2009s). Logistic regression and receiver operating characteristic curve analysis were performed to evaluate diagnostic performance and determine optimal cut\u2010off values. HS patients exhibited significantly altered hematologic parameters compared with controls, including higher reticulocyte counts, red cell distribution width, and mean corpuscular hemoglobin. The EMA\u2010binding test demonstrated high specificity (100%) but lower sensitivity (57.9%). OFT achieved high sensitivity (&gt;97%) but low specificity (&lt;47%). AGLT showed balanced accuracy (sensitivity 68.4%, specificity 96.1%). OFT\u2010FCM yielded areas under the curve of 0.85 for both protocols, with optimal thresholds providing specificities of 95\u2013100% and sensitivities of 57\u201359%. No significant differences were observed between OFT\u2010FCM and EMA performance. OFT\u2010FCM effectively discriminates HS patients from healthy controls and showed diagnostic performance comparable to EMA and favorable relative to classical OFT and AGLT in this cohort, while offering practical advantages in terms of workflow simplicity and turnaround time, and supporting its use as a complementary flow\u2010cytometric assay within the diagnostic work\u2010up of HS.<\/jats:p>","DOI":"10.1002\/cyto.b.70040","type":"journal-article","created":{"date-parts":[[2026,5,13]],"date-time":"2026-05-13T04:55:08Z","timestamp":1778648108000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Diagnostic performance of flow cytometric osmotic fragility testing in hereditary spherocytosis"],"prefix":"10.1002","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9499-8999","authenticated-orcid":false,"given":"Silvia","family":"Escribano Serrat","sequence":"first","affiliation":[{"name":"Hemostasis and Erythropathology Laboratory, Hematopathology, Department of Pathology Biomedical Diagnostic Center (CDB), Hospital Cl\u00ednic de Barcelona  Barcelona Spain"},{"name":"Institut d'Investigacions Biom\u00e8diques August Pi i Sunyer (IDIBAPS)  Barcelona Spain"},{"name":"Facultat de Medicina I Ci\u00e9nces de la Salut University of Barcelona  Barcelona Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Patricia","family":"Molina","sequence":"additional","affiliation":[{"name":"Hemostasis and Erythropathology Laboratory, Hematopathology, Department of Pathology Biomedical Diagnostic Center (CDB), Hospital Cl\u00ednic de Barcelona  Barcelona Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pilar","family":"G\u00f3mez","sequence":"additional","affiliation":[{"name":"Hemostasis and Erythropathology Laboratory, Hematopathology, Department of Pathology Biomedical Diagnostic Center (CDB), Hospital Cl\u00ednic de Barcelona  Barcelona Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mireia","family":"Garre","sequence":"additional","affiliation":[{"name":"Hemostasis and Erythropathology Laboratory, Hematopathology, Department of Pathology Biomedical Diagnostic Center (CDB), Hospital Cl\u00ednic de Barcelona  Barcelona Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Meritxell","family":"Nomdedeu","sequence":"additional","affiliation":[{"name":"Institut d'Investigacions Biom\u00e8diques August Pi i Sunyer (IDIBAPS)  Barcelona Spain"},{"name":"Facultat de Medicina I Ci\u00e9nces de la Salut University of Barcelona  Barcelona Spain"},{"name":"Hemotherapy and Hemostasis Department, Cancer and Blood Diseases Institute Hospital Cl\u00ednic de Barcelona  Barcelona Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Blanca","family":"De Moner","sequence":"additional","affiliation":[{"name":"Hemostasis and Erythropathology Laboratory, Hematopathology, Department of Pathology Biomedical Diagnostic Center (CDB), Hospital Cl\u00ednic de Barcelona  Barcelona Spain"},{"name":"Institut d'Investigacions Biom\u00e8diques August Pi i Sunyer (IDIBAPS)  Barcelona Spain"},{"name":"Facultat de Medicina I Ci\u00e9nces de la Salut University of Barcelona  Barcelona Spain"},{"name":"Josep Carreras Leukaemia Research Institute Hospital Cl\u00ednic de Barcelona  Barcelona Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Estefan\u00eda","family":"Garc\u00eda","sequence":"additional","affiliation":[{"name":"Hemostasis and Erythropathology Laboratory, Hematopathology, Department of Pathology Biomedical Diagnostic Center (CDB), Hospital Cl\u00ednic de Barcelona  Barcelona Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ana Bel\u00e9n","family":"Moreno\u2010Casta\u00f1o","sequence":"additional","affiliation":[{"name":"Hemostasis and Erythropathology Laboratory, Hematopathology, Department of Pathology Biomedical Diagnostic Center (CDB), Hospital Cl\u00ednic de Barcelona  Barcelona Spain"},{"name":"Institut d'Investigacions Biom\u00e8diques August Pi i Sunyer (IDIBAPS)  Barcelona Spain"},{"name":"Facultat de Medicina I Ci\u00e9nces de la Salut University of Barcelona  Barcelona Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"\u00c1lex","family":"Ramos","sequence":"additional","affiliation":[{"name":"Hemostasis and Erythropathology Laboratory, Hematopathology, Department of Pathology Biomedical Diagnostic Center (CDB), Hospital Cl\u00ednic de Barcelona  Barcelona Spain"},{"name":"Facultat de Medicina I Ci\u00e9nces de la Salut University of Barcelona  Barcelona Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Julia","family":"Martinez\u2010Sanchez","sequence":"additional","affiliation":[{"name":"Hemostasis and Erythropathology Laboratory, Hematopathology, Department of Pathology Biomedical Diagnostic Center (CDB), Hospital Cl\u00ednic de Barcelona  Barcelona Spain"},{"name":"Institut d'Investigacions Biom\u00e8diques August Pi i Sunyer (IDIBAPS)  Barcelona Spain"},{"name":"Facultat de Medicina I Ci\u00e9nces de la Salut University of Barcelona  Barcelona Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marc","family":"Pino","sequence":"additional","affiliation":[{"name":"Hemostasis and Erythropathology Laboratory, Hematopathology, Department of Pathology Biomedical Diagnostic Center (CDB), Hospital Cl\u00ednic de Barcelona  Barcelona Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gin\u00e9s","family":"Escolar","sequence":"additional","affiliation":[{"name":"Hemostasis and Erythropathology Laboratory, Hematopathology, Department of Pathology Biomedical Diagnostic Center (CDB), Hospital Cl\u00ednic de Barcelona  Barcelona Spain"},{"name":"Institut d'Investigacions Biom\u00e8diques August Pi i Sunyer (IDIBAPS)  Barcelona Spain"},{"name":"Facultat de Medicina I Ci\u00e9nces de la Salut University of Barcelona  Barcelona Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maribel","family":"D\u00edaz\u2010Ricart","sequence":"additional","affiliation":[{"name":"Hemostasis and Erythropathology Laboratory, Hematopathology, Department of Pathology Biomedical Diagnostic Center (CDB), Hospital Cl\u00ednic de Barcelona  Barcelona Spain"},{"name":"Institut d'Investigacions Biom\u00e8diques August Pi i Sunyer (IDIBAPS)  Barcelona Spain"},{"name":"Facultat de Medicina I Ci\u00e9nces de la Salut University of Barcelona  Barcelona Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2026,5,12]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/ijlh.12794"},{"key":"e_1_2_8_3_1","volume-title":"Dacie and Lewis practical haematology","author":"Bain B. J.","year":"2006"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.3324\/haematol.2011.052845"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.3324\/haematol.2012.075150"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2141.2011.08921.x"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2141.2004.05052.x"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.5045\/br.2022.2021224"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.b.21511"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1002\/pbc.24157"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00277-010-1112-0"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.blre.2013.04.003"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF01715351"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2141.2007.06944.x"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1002\/cytob.21187"},{"key":"e_1_2_8_16_1","unstructured":"ICSH. (n.d.).Guidelines for the laboratory diagnosis of nonimmune hereditary red cell membrane disorders.https:\/\/www.icsh.org\/guidelines-for-the-laboratory-diagnosis-of-nonimmune-hereditary-red-cell-membrane-disorders"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.3324\/haematol.2016.161166"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1751-553X.2008.01098.x"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.0141-9854.2003.00557.x"},{"issue":"3","key":"e_1_2_8_20_1","first-page":"924","article-title":"Rapid flow cytometric test for the diagnosis of membrane cytoskeleton\u2010associated haemolytic anaemia","volume":"111","author":"King M. J.","year":"2000","journal-title":"British Journal of Haematology"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1111\/ijlh.12070"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.3324\/haematol.12546"},{"key":"e_1_2_8_23_1","volume-title":"Gu\u00eda esferocitosis hereditaria. Diagn\u00f3stico y manejo en poblaci\u00f3n pedi\u00e1trica","author":"Mart\u00edn\u2010Consuegra S."},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.b.21205"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.b.21733"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1309\/AJCPO7V4OGXLIIPP"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(08)61588-3"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2141.1997.d01-2074.x"},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.1002\/cytob.21143"},{"key":"e_1_2_8_30_1","doi-asserted-by":"publisher","DOI":"10.1159\/000094679"},{"issue":"3","key":"e_1_2_8_31_1","first-page":"570","article-title":"Rapid flow cytometric test using eosin\u20105\u2010maleimide for diagnosis of red blood cell membrane disorders","volume":"40","author":"Tachavanich K.","year":"2009","journal-title":"The Southeast Asian Journal of Tropical Medicine and Public Health"},{"key":"e_1_2_8_32_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00277-014-2270-2"},{"key":"e_1_2_8_33_1","doi-asserted-by":"publisher","DOI":"10.1159\/000438738"},{"key":"e_1_2_8_34_1","volume-title":"Manual de t\u00e9cnicas de laboratorio en hematolog\u00eda","author":"Vives Corrons J. L.","year":"2014"},{"key":"e_1_2_8_35_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.b.20583"},{"key":"e_1_2_8_36_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.b.22254"},{"key":"e_1_2_8_37_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.b.20448"},{"key":"e_1_2_8_38_1","doi-asserted-by":"publisher","DOI":"10.1002\/jcla.24034"},{"key":"e_1_2_8_39_1","doi-asserted-by":"publisher","DOI":"10.1177\/2211068214532254"},{"key":"e_1_2_8_40_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2141.1980.tb07167.x"}],"container-title":["Cytometry Part B: Clinical Cytometry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cyto.b.70040","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/cyto.b.70040","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cyto.b.70040","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,13]],"date-time":"2026-05-13T04:55:27Z","timestamp":1778648127000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/cyto.b.70040"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,5,12]]},"references-count":39,"alternative-id":["10.1002\/cyto.b.70040"],"URL":"https:\/\/doi.org\/10.1002\/cyto.b.70040","archive":["Portico"],"relation":{},"ISSN":["1552-4949","1552-4957"],"issn-type":[{"value":"1552-4949","type":"print"},{"value":"1552-4957","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,5,12]]},"assertion":[{"value":"2026-02-15","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-05-05","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-05-12","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"cyto.b.70040"}}