{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,8,25]],"date-time":"2025-08-25T04:40:05Z","timestamp":1756096805616,"version":"3.44.0"},"reference-count":21,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,8,25]],"date-time":"2025-08-25T00:00:00Z","timestamp":1756080000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Vet. Sci."],"abstract":"<jats:p>Mast cell tumors (MCTs) are the most common skin neoplasms in dogs and exhibit highly variable biological behavior. Metastasis primarily affects the lymph nodes, though less frequently, MCTs can infiltrate the spleen, liver, peripheral blood, and bone marrow. Flow cytometry of fine needle aspirate samples represents a non-invasive diagnostic procedure that has shown promise for detecting and quantifying mast cells in primary tumors and lymph nodes. However, analytical validation of this method for clinical use is lacking. This study aimed to evaluate the analytical performance of a flow cytometric panel for quantifying mast cells in peripheral blood, bone marrow, and lymph node aspirates from dogs. Key parameters as the limit of blank (LOB), lower limit of detection (LLoD), lower limit of quantification (LLoQ), intra-assay precision, and accuracy, were evaluated. The method demonstrated high precision across a wide range of mast cell concentrations, with analytical coefficient of variation (CV<jats:sub>A<\/jats:sub>) of less than 10% for all sample types. It also showed good accuracy with minimal proportional bias observed in lymph node samples, particularly at higher mast cell concentrations. The LLoQ was 0.1% for all sample types. Flow cytometry provided reliable results highlighting its potential as a clinical tool for diagnosing and staging MCTs. These findings support the clinical applicability of flow cytometry as a minimally invasive, highly accurate method for assessing mast cell infiltration in peripheral blood, bone marrow, and lymph nodes, offering an alternative to traditional microscopic examination. This validation establishes a foundation for future studies on the prognostic implications of mast cell infiltration in MCT progression.<\/jats:p>","DOI":"10.3389\/fvets.2025.1542460","type":"journal-article","created":{"date-parts":[[2025,8,25]],"date-time":"2025-08-25T04:10:25Z","timestamp":1756095025000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Analytical validation of a flow cytometric method for the detection and quantification of canine mast cells in peripheral blood, bone marrow, and lymph node"],"prefix":"10.3389","volume":"12","author":[{"given":"Giulia","family":"Iamone","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fulvio","family":"Riondato","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rachel","family":"Hanford","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Amandine","family":"Lejeune","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Amir","family":"Kol","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2025,8,25]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","DOI":"10.3390\/cells11040618","article-title":"Diagnosis, prognosis and treatment of canine cutaneous and subcutaneous mast cell tumors","volume":"11","author":"de Nardi","year":"2022","journal-title":"Cells"},{"key":"ref2","doi-asserted-by":"publisher","first-page":"141","DOI":"10.2478\/jvetres-2019-0002","article-title":"Occurrence and distribution of canine cutaneous mast cell tumour characteristics among predisposed breeds","volume":"63","author":"\u015amiech","year":"2019","journal-title":"J Vet Res (Poland)"},{"key":"ref3","doi-asserted-by":"publisher","first-page":"469","DOI":"10.1177\/030098588402100503","article-title":"Canine cutaneous mast cell tumor: morphologic grading and survival time in 83 dogs","volume":"21","author":"Patnaik","year":"1984","journal-title":"Vet Pathol"},{"key":"ref4","doi-asserted-by":"publisher","first-page":"147","DOI":"10.1177\/0300985810386469","article-title":"Proposal of a 2-tier histologic grading system for canine cutaneous mast cell tumors to more accurately predict biological behavior","volume":"48","author":"Kiupel","year":"2011","journal-title":"Vet Pathol"},{"key":"ref5","doi-asserted-by":"publisher","first-page":"81","DOI":"10.1111\/j.1476-5829.2009.00183.x","article-title":"A review of sentinel lymph node evaluation and the need for its incorporation into veterinary oncology","volume":"7","author":"Tuohy","year":"2009","journal-title":"Vet Comp Oncol"},{"key":"ref6","doi-asserted-by":"publisher","first-page":"329","DOI":"10.1016\/j.jcpa.2014.07.004","article-title":"Correlation of nodal mast cells with clinical outcome in dogs with mast cell tumour and a proposed classification system for the evaluation of node metastasis","volume":"151","author":"Weishaar","year":"2014","journal-title":"J Comp Pathol"},{"key":"ref7","doi-asserted-by":"publisher","first-page":"1206","DOI":"10.1111\/vco.12256","article-title":"Cytologic\u2013histologic concordance in the diagnosis of neoplasia in canine and feline lymph nodes: a retrospective study of 367 cases","volume":"15","author":"Ku","year":"2017","journal-title":"Vet Comp Oncol"},{"key":"ref8","doi-asserted-by":"publisher","first-page":"15","DOI":"10.1016\/j.tvjl.2018.10.004","article-title":"Utility of flow cytometry in canine primary cutaneous and matched nodal mast cell tumor","volume":"242","author":"Sulce","year":"2018","journal-title":"Vet J"},{"key":"ref9","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1111\/vco.13019","article-title":"Flow cytometry for the detection and quantification of mast cells in lymph nodes: a prospective study in 64 dogs with mast cell tumour","volume":"23","author":"Iamone","year":"2024","journal-title":"Vet Comp Oncol"},{"key":"ref10","doi-asserted-by":"publisher","first-page":"1648","DOI":"10.2460\/javma.1992.200.11.1648","article-title":"Determination of the number of mast cells in lymph node, bone marrow, and buffy coat cytologic specimens from dogs","volume":"200","author":"Bookbinder","year":"1992","journal-title":"J Am Vet Med Assoc"},{"key":"ref11","doi-asserted-by":"publisher","first-page":"e988","DOI":"10.1002\/cpz1.988","article-title":"Flow cytometry method validation protocols","volume":"4","author":"Selliah","year":"2024","journal-title":"Curr Protoc"},{"key":"ref12","doi-asserted-by":"publisher","first-page":"553","DOI":"10.1111\/vco.12520","article-title":"Effects of ibrutinib on proliferation and histamine release in canine neoplastic mast cells","volume":"17","author":"Gamperl","year":"2019","journal-title":"Vet Comp Oncol"},{"key":"ref13","doi-asserted-by":"publisher","first-page":"1016","DOI":"10.1038\/leu.2017.338","article-title":"Drug-induced inhibition of phosphorylation of STAT5 overrides drug resistance in neoplastic mast cells","volume":"32","author":"Peter","year":"2018","journal-title":"Leukemia"},{"key":"ref14","doi-asserted-by":"publisher","first-page":"1240","DOI":"10.1111\/vco.12260","article-title":"IL-4 downregulates expression of the target receptor CD30 in neoplastic canine mast cells","volume":"15","author":"Bauer","year":"2017","journal-title":"Vet Comp Oncol"},{"key":"ref15","doi-asserted-by":"publisher","first-page":"10","DOI":"10.3389\/fvets.2023.1279881","article-title":"Development of a flow cytometric panel to assess prognostic biomarkers in fine needle aspirates of canine cutaneous or subcutaneous mast cell tumors","volume":"10","author":"Wu","year":"2023","journal-title":"Front Vet Sci"},{"key":"ref16","doi-asserted-by":"publisher","first-page":"858","DOI":"10.1111\/j.1398-9995.2012.02833.x","article-title":"NI-1: a novel canine mastocytoma model for studying drug resistance and IgER-dependent mast cell activation","volume":"67","author":"Hadzijusufovic","year":"2012","journal-title":"Allergy"},{"key":"ref17","doi-asserted-by":"publisher","first-page":"259","DOI":"10.1111\/vcp.12735","article-title":"Reporting results with (un)certainty","volume":"48","author":"Moore","year":"2019","journal-title":"Vet Clin Pathol"},{"key":"ref18","article-title":"Mast cell- and eosinophil-mediated hypersensitivity","author":"Tizard","year":"","journal-title":"Veterinary Immunology"},{"key":"ref19","doi-asserted-by":"publisher","first-page":"5","DOI":"10.1016\/j.jcpa.2018.10.170","article-title":"Chronic basophilic Leukaemia in a dog","volume":"166","author":"Azakami","year":"2019","journal-title":"J Comp Pathol"},{"key":"ref20","doi-asserted-by":"publisher","first-page":"813","DOI":"10.5858\/2008-132-813-ISOBBM","article-title":"Immunophenotypic study of basophils by multiparameter flow cytometry","volume":"132","author":"Han","year":"2008","journal-title":"Arch Pathol Lab Med"},{"key":"ref21","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1016\/B978-0-323-53314-0.00004-3","article-title":"Round cells","volume-title":"Cowell and Tyler\u2019s Diagnostic Cytology and Hematology of the Dog and Cat","author":"Jacocks","year":"2020"}],"container-title":["Frontiers in Veterinary Science"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fvets.2025.1542460\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,8,25]],"date-time":"2025-08-25T04:10:26Z","timestamp":1756095026000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fvets.2025.1542460\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,8,25]]},"references-count":21,"alternative-id":["10.3389\/fvets.2025.1542460"],"URL":"https:\/\/doi.org\/10.3389\/fvets.2025.1542460","relation":{},"ISSN":["2297-1769"],"issn-type":[{"value":"2297-1769","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,8,25]]},"article-number":"1542460"}}