{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T21:37:28Z","timestamp":1789767448001,"version":"4.0.1"},"reference-count":71,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2021,9,30]],"date-time":"2021-09-30T00:00:00Z","timestamp":1632960000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico, Bras\u00edlia, Brazil (CNPQ), Bras\u00edlia, Brazi","award":["303765\/2018-6, \t409440\/2016-7, 400194\/2014-7"],"award-info":[{"award-number":["303765\/2018-6, \t409440\/2016-7, 400194\/2014-7"]}]},{"name":"Funda\u00e7\u00e3o de Amparo \u00e0 Pesquisa do Estado do Rio de Janeiro, Brazil (FAPERJ)","award":["E26\/110.105\/2014, E-26\/010.101259\/2018 and E26\/102.191\/2013"],"award-info":[{"award-number":["E26\/110.105\/2014, E-26\/010.101259\/2018 and E26\/102.191\/2013"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancers"],"abstract":"<jats:p>Early diagnosis of pediatric cancer is key for adequate patient management and improved outcome. Although multiparameter flow cytometry (MFC) has proven of great utility in the diagnosis and classification of hematologic malignancies, its application to non-hematopoietic pediatric tumors remains limited. Here we designed and prospectively validated a new single eight-color antibody combination\u2014solid tumor orientation tube, STOT\u2014for diagnostic screening of pediatric cancer by MFC. A total of 476 samples (139 tumor mass, 138 bone marrow, 86 lymph node, 58 peripheral blood, and 55 other body fluid samples) from 296 patients with diagnostic suspicion of pediatric cancer were analyzed by MFC vs. conventional diagnostic procedures. STOT was designed after several design\u2013test\u2013evaluate\u2013redesign cycles based on a large panel of monoclonal antibody combinations tested on 301 samples. In its final version, STOT consists of a single 8-color\/12-marker antibody combination (CD99-CD8\/numyogenin\/CD4-EpCAM\/CD56\/GD2\/smCD3-CD19\/cyCD3-CD271\/CD45). Prospective validation of STOT in 149 samples showed concordant results with the patient WHO\/ICCC-3 diagnosis in 138\/149 cases (92.6%). These included: 63\/63 (100%) reactive\/disease-free samples, 43\/44 (98%) malignant and 4\/4 (100%) benign non-hematopoietic tumors together with 28\/38 (74%) leukemia\/lymphoma cases; the only exception was Hodgkin lymphoma that required additional markers to be stained. In addition, STOT allowed accurate discrimination among the four most common subtypes of malignant CD45\u2212 CD56++ non-hematopoietic solid tumors: 13\/13 (GD2++\u00a0numyogenin\u2212 CD271\u2212\/+\u00a0nuMyoD1\u2212 CD99\u2212 EpCAM\u2212) neuroblastoma samples, 5\/5 (GD2\u2212\u00a0numyogenin++ CD271++\u00a0nuMyoD1++ CD99\u2212\/+ EpCAM\u2212) rhabdomyosarcomas, 2\/2 (GD2\u2212\/+\u00a0numyogenin\u2212 CD271+\u00a0nuMyoD1\u2212 CD99+ EpCAM\u2212) Ewing sarcoma family of tumors, and 7\/7 (GD2\u2212\u00a0numyogenin\u2212 CD271+\u00a0nuMyoD1\u2212 CD99\u2212 EpCAM+) Wilms tumors. In summary, here we designed and validated a new standardized antibody combination and MFC assay for diagnostic screening of pediatric solid tumors that might contribute to fast and accurate diagnostic orientation and classification of pediatric cancer in routine clinical practice.<\/jats:p>","DOI":"10.3390\/cancers13194945","type":"journal-article","created":{"date-parts":[[2021,10,1]],"date-time":"2021-10-01T10:55:40Z","timestamp":1633085740000},"page":"4945","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Flow Cytometry Immunophenotyping for Diagnostic Orientation and Classification of Pediatric Cancer Based on the EuroFlow Solid Tumor Orientation Tube (STOT)"],"prefix":"10.3390","volume":"13","author":[{"given":"Cristiane de S\u00e1","family":"Ferreira-Facio","sequence":"first","affiliation":[{"name":"Internal Medicine Postgraduate Program, Faculty of Medicine, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro 21941-617, Brazil"},{"name":"Cytometry Service, Institute of Paediatrics and Puericultura Martag\u00e3o Gesteira (IPPMG), Faculty of Medicine, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro 21941-612, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8513-7003","authenticated-orcid":false,"given":"Vitor","family":"Botafogo","sequence":"additional","affiliation":[{"name":"Internal Medicine Postgraduate Program, Faculty of Medicine, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro 21941-617, Brazil"},{"name":"Cytometry Service, Institute of Paediatrics and Puericultura Martag\u00e3o Gesteira (IPPMG), Faculty of Medicine, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro 21941-612, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0680-8372","authenticated-orcid":false,"given":"Patr\u00edcia Mello","family":"Ferr\u00e3o","sequence":"additional","affiliation":[{"name":"Cytometry Service, Institute of Paediatrics and Puericultura Martag\u00e3o Gesteira (IPPMG), Faculty of Medicine, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro 21941-612, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria Clara","family":"Canellas","sequence":"additional","affiliation":[{"name":"Cytometry Service, Institute of Paediatrics and Puericultura Martag\u00e3o Gesteira (IPPMG), Faculty of Medicine, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro 21941-612, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cristiane B.","family":"Milito","sequence":"additional","affiliation":[{"name":"Department of Pathology, Faculty of Medicine, Clementino Fraga Filho University Hospital, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro 21941-617, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S\u00e9rgio","family":"Romano","sequence":"additional","affiliation":[{"name":"Laboratory of Anatomical Pathology and Cytopathology, Instituto Nacional de C\u00e2ncer (INCa), Rio de Janeiro 20220-400, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1936-1952","authenticated-orcid":false,"given":"Daiana V.","family":"Lopes","sequence":"additional","affiliation":[{"name":"Cytometry Service, Institute of Paediatrics and Puericultura Martag\u00e3o Gesteira (IPPMG), Faculty of Medicine, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro 21941-612, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lisandra C.","family":"Teixeira","sequence":"additional","affiliation":[{"name":"Internal Medicine Postgraduate Program, Faculty of Medicine, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro 21941-617, Brazil"},{"name":"Cytometry Service, Institute of Paediatrics and Puericultura Martag\u00e3o Gesteira (IPPMG), Faculty of Medicine, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro 21941-612, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elen","family":"Oliveira","sequence":"additional","affiliation":[{"name":"Internal Medicine Postgraduate Program, Faculty of Medicine, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro 21941-617, Brazil"},{"name":"Cytometry Service, Institute of Paediatrics and Puericultura Martag\u00e3o Gesteira (IPPMG), Faculty of Medicine, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro 21941-612, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Enrico","family":"Bruno-Riscarolli","sequence":"additional","affiliation":[{"name":"Internal Medicine Postgraduate Program, Faculty of Medicine, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro 21941-617, Brazil"},{"name":"Cytometry Service, Institute of Paediatrics and Puericultura Martag\u00e3o Gesteira (IPPMG), Faculty of Medicine, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro 21941-612, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fabiana V.","family":"Mello","sequence":"additional","affiliation":[{"name":"Cytometry Service, Institute of Paediatrics and Puericultura Martag\u00e3o Gesteira (IPPMG), Faculty of Medicine, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro 21941-612, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patr\u00edcia F. R.","family":"Siqueira","sequence":"additional","affiliation":[{"name":"Internal Medicine Postgraduate Program, Faculty of Medicine, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro 21941-617, Brazil"},{"name":"Cytometry Service, Institute of Paediatrics and Puericultura Martag\u00e3o Gesteira (IPPMG), Faculty of Medicine, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro 21941-612, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patr\u00edcia","family":"Moura","sequence":"additional","affiliation":[{"name":"I\u2019Dor Institute, Hospital Estadual da Crian\u00e7a, Rio de Janeiro 21330-400, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francisco Nicanor","family":"Macedo","sequence":"additional","affiliation":[{"name":"I\u2019Dor Institute, Hospital Estadual da Crian\u00e7a, Rio de Janeiro 21330-400, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Danielle N.","family":"Forny","sequence":"additional","affiliation":[{"name":"Department of Pediatric Surgery, Institute of Paediatrics and Puericultura Martag\u00e3o Gesteira (IPPMG), Faculty of Medicine, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro 21941-612, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lu\u00edza","family":"Simi\u00e3o","sequence":"additional","affiliation":[{"name":"Cytometry Service, Institute of Paediatrics and Puericultura Martag\u00e3o Gesteira (IPPMG), Faculty of Medicine, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro 21941-612, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana Lu\u00edza","family":"Pureza","sequence":"additional","affiliation":[{"name":"Cytometry Service, Institute of Paediatrics and Puericultura Martag\u00e3o Gesteira (IPPMG), Faculty of Medicine, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro 21941-612, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marcelo Gerardin Poirot","family":"Land","sequence":"additional","affiliation":[{"name":"Internal Medicine Postgraduate Program, Faculty of Medicine, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro 21941-617, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9312-4023","authenticated-orcid":false,"given":"Carlos Eduardo","family":"Pedreira","sequence":"additional","affiliation":[{"name":"Systems and Computing Engineering Department (COPPE-PESC), Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro 21941-972, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3650-7087","authenticated-orcid":false,"given":"Jacques J. M. van","family":"Dongen","sequence":"additional","affiliation":[{"name":"Department of Immunohematology and Blood Transfusion (IHB), Leiden University Medical Center (LUMC), 2333 ZA Leiden, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0007-7230","authenticated-orcid":false,"given":"Alberto","family":"Orfao","sequence":"additional","affiliation":[{"name":"Translational and Clinical Research Program, Centro de Investigaci\u00f3n del C\u00e1ncer and IBMCC (CSIC-University of Salamanca), Cytometry Service, NUCLEUS, Department of Medicine, University of Salamanca (USAL), Institute of Biomedical Research of Salamanca (IBSAL), 37007 Salamanca, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5340-5816","authenticated-orcid":false,"given":"Elaine Sobral da","family":"Costa","sequence":"additional","affiliation":[{"name":"Internal Medicine Postgraduate Program, Faculty of Medicine, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro 21941-617, Brazil"},{"name":"Cytometry Service, Institute of Paediatrics and Puericultura Martag\u00e3o Gesteira (IPPMG), Faculty of Medicine, Federal University of Rio de Janeiro (UFRJ), Rio de Janeiro 21941-612, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,9,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"483","DOI":"10.1016\/S1470-2045(18)30909-4","article-title":"Estimating the total incidence of global childhood cancer: A simulation-based analysis","volume":"20","author":"Ward","year":"2019","journal-title":"Lancet Oncol."},{"key":"ref_2","unstructured":"Triche, T.J., Hicks, J., and Sorensen, P.H.B. 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