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Flow cytometry plays a critical role in the diagnosis and management of the disease. It is used to establish the abnormal immature T\u2010cell phenotype and to distinguish the early T\u2010cell precursor (ETP)\u2010ALL from more mature types at diagnosis. The evaluation of mediastinal disease is often complicated by the difficulty of the phenotypic distinction between the normal thymic precursors and the abnormal T lymphoblasts. Follow\u2010up measurements of minimal\/measurable residual disease (MRD) are critical for therapy decision\u2010making and prognostication. In the MRD setting, flow cytometry requires a high degree of analytical expertise and assessment of numerous antigens. To address the diagnostic and monitoring challenges, we developed a single\u2010tube 21\u2010antigen assessment with simplified analysis. The assay distinguishes between normal thymic precursors and T lymphoblasts in tissue samples, enables evaluation of ETP versus non\u2010ETP phenotypes, and allows for MRD assessment below 0.01% robust to antigenic changes.<\/jats:p>","DOI":"10.1002\/cyto.b.22242","type":"journal-article","created":{"date-parts":[[2025,7,3]],"date-time":"2025-07-03T04:46:12Z","timestamp":1751517972000},"page":"299-311","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["A 20\u2010color 21\u2010antigen flow cytometric assay for disease monitoring of T\u2010cell lymphoblastic leukemia"],"prefix":"10.1002","volume":"108","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1054-4418","authenticated-orcid":false,"given":"Qi","family":"Gao","sequence":"first","affiliation":[{"name":"Department of Pathology and Laboratory Medicine Memorial Sloan Kettering Cancer Center  New York New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingping","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine Memorial Sloan Kettering Cancer Center  New York New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Krasimira","family":"Rozenova","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine Memorial Sloan Kettering Cancer Center  New York New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaotian","family":"Sun","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine Memorial Sloan Kettering Cancer Center  New York New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amanda","family":"Burke","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine Memorial Sloan Kettering Cancer Center  New York New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Olivia","family":"Miu","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine Memorial Sloan Kettering Cancer Center  New York New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nghia","family":"Nguyen","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine Memorial Sloan Kettering Cancer Center  New York New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shu Jie","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine Memorial Sloan Kettering Cancer Center  New York New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mikhail","family":"Roshal","sequence":"additional","affiliation":[{"name":"Department of Pathology and Laboratory Medicine Memorial Sloan Kettering Cancer Center  New York New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,7,3]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2016-03-643544"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood.2022015850"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature06840"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrc.2016.63"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1001\/jamaoncol.2017.0580"},{"key":"e_1_2_6_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.leu.2402736"},{"key":"e_1_2_6_8_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2008-01-132837"},{"key":"e_1_2_6_9_1","doi-asserted-by":"publisher","DOI":"10.1200\/JCO.2005.02.2707"},{"key":"e_1_2_6_10_1","doi-asserted-by":"publisher","DOI":"10.1002\/cpz1.884"},{"key":"e_1_2_6_11_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood.2022015646"},{"key":"e_1_2_6_12_1","unstructured":"CLSI. 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