{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T19:23:54Z","timestamp":1790796234381,"version":"4.1.0"},"reference-count":34,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2020,9,8]],"date-time":"2020-09-08T00:00:00Z","timestamp":1599523200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100007052","name":"Memorial Sloan Kettering Cancer Center","doi-asserted-by":"crossref","award":["P30 CA008748"],"award-info":[{"award-number":["P30 CA008748"]}],"id":[{"id":"10.13039\/100007052","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Cytometry Part B Clinical"],"published-print":{"date-parts":[[2021,7]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Introduction<\/jats:title><jats:p>The diagnosis of CLL\/SLL relies on flow cytometric immunophenotyping. Increasing emphasis is being placed on precise detection of the minimal residual disease. Following antigen recommendations of ERIC and ESCCA's Harmonization Project, we validated a 14\u2010color assay for the characterization CD5+ lymphoproliferative neoplasms and CLL MRD with a sensitivity of at least 10<jats:sup>\u22124<\/jats:sup>.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>The assay was designed based on ERIC\/ESCCA recommended antigens with the addition of CD40 for alternate gating when CD19 expression is reduced. Lower limit of quantitation\/lower limit of detection, assay procedural precision, linearity, and limit of blank were established. Then, 52 CD5+ B\u2010cell lymphoproliferative neoplasms (41 CLL\/11 non\u2010CLL) and 29 normal samples were used for parallel evaluation. Automated cluster identification and quantitation of CLL clones in MRD setting was performed using Barned\u2010Hutt SNE. Separation analysis between CLL and non\u2010CLL phenotypes was performed by PCA and bh\u2010SNE.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Separation ratios for each antigen exceeded ERIC\/ESCCA guidelines. Precision was &lt;20% at LLOQ (0.01%). The limit of blank was &lt;10\/500,000 cells. Concordance between the 14\u2010color and legacy assay (Deming regression <jats:italic>y<\/jats:italic>\u00a0=\u00a01.01x, <jats:italic>r<\/jats:italic><jats:sup>2<\/jats:sup>\u00a0=\u00a0.99) was seen. All 20 samples with MRD levels 0.5%\u20130.006% (median 0.04%) showed an abnormal cell cluster by bh\u2010SNE, with concordant results between manual and automated quantitation (<jats:italic>y<\/jats:italic>\u00a0=\u00a0x, <jats:italic>r<\/jats:italic><jats:sup>2<\/jats:sup>\u00a0=\u00a01). CLL cases clustered together and away from mantle cell lymphoma by bh\u2010SNE and PCA with outlier atypical phenotype CLL cases posing diagnostic challenges by both manual and automated analysis.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>The 14\u2010color CD5+ LPD assay provides a robust standardization platform for MRD and disease characterization using both manual and automated analysis.<\/jats:p><\/jats:sec>","DOI":"10.1002\/cyto.b.21953","type":"journal-article","created":{"date-parts":[[2020,9,14]],"date-time":"2020-09-14T07:45:22Z","timestamp":1600069522000},"page":"509-518","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["14\u2010Color single tube for flow cytometric characterization of CD5+ B\u2010LPDs and high sensitivity automated minimal residual disease quantitation of CLL\/SLL"],"prefix":"10.1002","volume":"100","author":[{"given":"Jennifer M.","family":"Goshaw","sequence":"first","affiliation":[{"name":"Memorial Sloan Kettering Cancer Center, Department of Pathology Hematopathology Service  New York New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qi","family":"Gao","sequence":"additional","affiliation":[{"name":"Memorial Sloan Kettering Cancer Center, Department of Pathology Hematopathology Service  New York New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica","family":"Wardrope","sequence":"additional","affiliation":[{"name":"Memorial Sloan Kettering Cancer Center, Department of Pathology Hematopathology Service  New York New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ahmet","family":"Dogan","sequence":"additional","affiliation":[{"name":"Memorial Sloan Kettering Cancer Center, Department of Pathology Hematopathology Service  New York New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mikhail","family":"Roshal","sequence":"additional","affiliation":[{"name":"Memorial Sloan Kettering Cancer Center, Department of Pathology Hematopathology Service  New York New York USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2020,9,8]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.2594"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/leu.2017.274"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/leu.2010.160"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.3324\/haematol.2017.183954"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1002\/cyto.b.21252"},{"issue":"2","key":"e_1_2_7_7_1","first-page":"103","article-title":"Optimization and validation of an 8\u2010color single\u2010tube assay for the sensitive detection of minimal residual disease in B\u2010cell chronic lymphocytic leukemia detected via flow cytometry","volume":"47","author":"Dowling A. 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