{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,19]],"date-time":"2026-10-19T16:01:12Z","timestamp":1792425672566,"version":"4.5.0"},"reference-count":48,"publisher":"Wiley","issue":"8","license":[{"start":{"date-parts":[[2025,7,9]],"date-time":"2025-07-09T00:00:00Z","timestamp":1752019200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Cytometry Pt A"],"published-print":{"date-parts":[[2025,8]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>Systemic mastocytosis (SM) is a neoplastic disease characterized by abnormal mast cell (MC) activation and proliferation. Accurate diagnosis often relies on flow cytometry to detect aberrant CD25, CD2, and CD30 expression on MCs in bone marrow (BM). However, the frequently low abundance of MCs in BM, lack of completely specific antigens, and strong and highly variable autofluorescence can cause misinterpretation and lead to diagnostic misclassifications. We investigated the potentially interfering cell populations in flow cytometric analysis of MCs based on literature and expert insights, focusing on CD117, CD45, CD203c, and Fc\u03b5R1. Additionally, we determined the most appropriate approach to quantify aberrant CD25, CD2, and CD30 expression. Apoptotic granulocytes frequently cause misinterpretation by mimicking strong CD117 and aberrant CD25, CD2, and CD30 expression, and must be distinguished from MCs with a viability dye like DRAQ7. CD117\u2010positive myeloblasts and promyelocytes overlap with CD117\u2010reduced immature MCs in advanced SM disease and can be differentiated using CD203c. Quantifying CD25, CD2, and CD30 expression is skewed on log\u2010transformed scales due to the strong and highly heterogeneous autofluorescence of MCs. Linear calculation of net expression levels of CD25, CD2, and CD30 yields the highest accuracies in predicting SM with a Youden index of 0.96, 0.93, and 0.88, respectively. Incorporating a viability dye like DRAQ7 and CD203c into the flow cytometric analysis for MC identification, along with the linear quantification of aberrant expression, significantly enhances the correct identification of MCs and increases the diagnostic accuracy of aberrant CD25, CD2, and CD30 expression for SM.<\/jats:p>","DOI":"10.1002\/cyto.a.24950","type":"journal-article","created":{"date-parts":[[2025,7,10]],"date-time":"2025-07-10T05:44:01Z","timestamp":1752126241000},"page":"538-550","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Critical Pitfalls in the Flow Cytometric Analysis of Mast Cells in Patients With Systemic Mastocytosis"],"prefix":"10.1002","volume":"107","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0708-8987","authenticated-orcid":false,"given":"Abdulrazzaq","family":"Alheraky","sequence":"first","affiliation":[{"name":"Department of Laboratory Medicine University Medical Center Groningen, University of Groningen  Groningen the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kees","family":"Meijer","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine University Medical Center Groningen, University of Groningen  Groningen the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marije T.","family":"Nijk","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine University Medical Center Groningen, University of Groningen  Groningen the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Saskia K.","family":"Klein","sequence":"additional","affiliation":[{"name":"Department of Hematology University Medical Center Groningen, University of Groningen  Groningen the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hanneke N. G.","family":"Oude Elberink","sequence":"additional","affiliation":[{"name":"Department of Allergology University of Groningen  Groningen the Netherlands"},{"name":"Groningen Research Institute for Asthma and COPD (GRIAC), University Medical Center Groningen University of Groningen  Groningen the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ido P.","family":"Kema","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine University Medical Center Groningen, University of Groningen  Groningen the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andr\u00e9 B.","family":"Mulder","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine University Medical Center Groningen, University of Groningen  Groningen the Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,7,9]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/nm.2755"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.iac.2006.05.006"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/ajh.26118"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.iac.2023.04.011"},{"key":"e_1_2_8_6_1","volume-title":"Haematolymphoid Tumours WHO, Part A, Classification of Tumours, 5th Edition","author":"WHO Classification of Tumours Editorial Board","year":"2024"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood.2022015850"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1182\/bloodadvances.2023009826"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-0716-1001-5_14"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.b.10072"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1189\/jlb.5RU0614-296R"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.iac.2014.01.008"},{"key":"e_1_2_8_13_1","first-page":"1493","article-title":"Flow Cytometric Analysis of Mast Cells From Normal and Pathological Human Bone Marrow Samples: Identification and Enumeration","volume":"149","author":"Orfao A.","year":"1996","journal-title":"American Journal of Pathology"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1309\/AJCPZ3J4GEEYIRRA"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1002\/clt2.12174"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaci.2014.05.047"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms222011270"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaci.2008.11.018"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1038\/leu.2012.120"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1182\/bloodadvances.2021005018"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ccell.2018.08.014"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.3109\/10428199809057558"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20052507"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2024-207798"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1309\/AJCP5PJWK4QFHWHM"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms21239030"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.iac.2006.05.008"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaci.2009.10.020"},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2012-07-442400"},{"key":"e_1_2_8_30_1","doi-asserted-by":"publisher","DOI":"10.1136\/jcp.2003.012864"},{"key":"e_1_2_8_31_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12032-012-0261-5"},{"issue":"9","key":"e_1_2_8_32_1","first-page":"2048","article-title":"Systemic Mastocytosis: Current Status and Challenges in 2024","volume":"9","author":"Ustun C.","year":"2025","journal-title":"Blood Advances"},{"key":"e_1_2_8_33_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaip.2021.02.040"},{"key":"e_1_2_8_34_1","doi-asserted-by":"publisher","DOI":"10.1159\/000329218"},{"key":"e_1_2_8_35_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2362.2007.01807.x"},{"key":"e_1_2_8_36_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood.2024024264"},{"issue":"1","key":"e_1_2_8_37_1","article-title":"No Adverse Effect of Residual Neoplastic Mast Cells in Systemic Mastocytosis Associated With Acute Myeloid Leukemia With T(8;21) (Q22;Q22); Runx1\u2010Runx1t1 After Bone Marrow Transplantation","volume":"2","author":"Dong Z. M. D.","year":"2014","journal-title":"Annals of Clinical Pathology"},{"key":"e_1_2_8_38_1","doi-asserted-by":"publisher","DOI":"10.1002\/0471142956.cy0941s65"},{"key":"e_1_2_8_39_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4939-7187-9_8"},{"key":"e_1_2_8_40_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1537-2995.1999.39090983.x"},{"key":"e_1_2_8_41_1","doi-asserted-by":"publisher","DOI":"10.1101\/pdb.prot087288"},{"key":"e_1_2_8_42_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-60327-017-5_2"},{"key":"e_1_2_8_43_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-30623-2"},{"key":"e_1_2_8_44_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.leukres.2003.10.035"},{"key":"e_1_2_8_45_1","doi-asserted-by":"publisher","DOI":"10.1038\/modpathol.2011.192"},{"key":"e_1_2_8_46_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2141.2003.04125.x"},{"key":"e_1_2_8_47_1","doi-asserted-by":"publisher","DOI":"10.1038\/modpathol.2010.224"},{"key":"e_1_2_8_48_1","doi-asserted-by":"publisher","DOI":"10.1111\/his.12221"},{"key":"e_1_2_8_49_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41375-024-02504-3"}],"container-title":["Cytometry Part A"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cyto.a.24950","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,9,9]],"date-time":"2025-09-09T14:32:17Z","timestamp":1757428337000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/cyto.a.24950"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,9]]},"references-count":48,"journal-issue":{"issue":"8","published-print":{"date-parts":[[2025,8]]}},"alternative-id":["10.1002\/cyto.a.24950"],"URL":"https:\/\/doi.org\/10.1002\/cyto.a.24950","archive":["Portico"],"relation":{},"ISSN":["1552-4922","1552-4930"],"issn-type":[{"value":"1552-4922","type":"print"},{"value":"1552-4930","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7,9]]},"assertion":[{"value":"2025-02-20","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-06-26","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-07-09","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}