{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T04:28:09Z","timestamp":1789705689893,"version":"4.0.1"},"reference-count":26,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,4,25]],"date-time":"2025-04-25T00:00:00Z","timestamp":1745539200000},"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:sec><jats:title>Introduction<\/jats:title><jats:p>The demand for non-invasive tumor biomarkers in veterinary field has recently grown significantly. Thymidine kinase 1 (TK1) is one of the non-invasive proliferation biomarkers that has been used for diagnosis and treatment monitoring of different canine malignancies. However, recent studies showed that the combination of TK1 with inflammatory biomarkers such as canine C-reactive protein (cCRP) can enhance the sensitivity for early tumor detection. Herein, we developed a machine learning (ML) model, i.e., Alertix-Cancer Risk Index (Alertix-CRI) which incorporates canine TK1 protein, CRP levels in conjunction with an age factor.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>A total of 287 serum samples were included in this study, consisting of 67 healthy dogs and dogs with different tumors (i.e., T-cell lymphoma <jats:italic>n<\/jats:italic> =\u202f24, B-cell lymphoma <jats:italic>n<\/jats:italic> =\u202f29, histiocytic sarcoma <jats:italic>n<\/jats:italic> =\u202f47, hemangiosarcoma <jats:italic>n<\/jats:italic> =\u202f26, osteosarcoma <jats:italic>n<\/jats:italic> =\u202f26, mastocytoma <jats:italic>n<\/jats:italic> =\u202f40, and mammary tumors <jats:italic>n<\/jats:italic> =\u202f28). Serum TK1 protein levels were measured using TK1-ELISA and cCRP levels by a quantitative ELISA. The whole data set was divided as training (70%) and validation (30%). The Alertix-Cancer Risk Index (Alertix-CRI) is a generalized boosted regression model (GBM) with high accuracy in the training set and further validation was carried out with the same model.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Both the TK1-ELISA and cCRP levels were significantly higher in the tumor group compared to healthy controls (<jats:italic>p<\/jats:italic>\u202f&amp;lt;\u202f0.0001). For overall tumors, the ROC curve analysis showed that TK1-ELISA has similar sensitivity as cCRP (54% vs. 51%) at a specificity of 95%. However, the Alertix-CRI for all malignancies showed an area under the curve (AUC) of 0.98, demonstrating very high discriminatory capacity, with a sensitivity of 90% and a specificity of 97%.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>These results demonstrate that the novel Alertix-CRI could be used as a decision-support tool helping clinicians to early differentiate dogs with malignant diseases from healthy. Additionally, these findings would facilitate the advancement of more precise and dependable diagnostic tools for early cancer detection and therapy monitoring within the realm of veterinary medicine.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fvets.2025.1570106","type":"journal-article","created":{"date-parts":[[2025,4,25]],"date-time":"2025-04-25T05:26:24Z","timestamp":1745558784000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":8,"title":["A novel cross-validated machine learning based Alertix-Cancer Risk Index for early detection of canine malignancies"],"prefix":"10.3389","volume":"12","author":[{"given":"Hanan","family":"Sharif","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Reza","family":"Arabi Belaghi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kiran Kumar","family":"Jagarlamudi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sara","family":"Saellstr\u00f6m","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liya","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Henrik","family":"R\u00f6nnberg","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Staffan","family":"Eriksson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,4,25]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"140","DOI":"10.1016\/j.molonc.2012.01.010","article-title":"Cancer biomarkers","volume":"6","author":"Henry","year":"2012","journal-title":"Mol Oncol"},{"key":"ref2","doi-asserted-by":"publisher","first-page":"1035","DOI":"10.2217\/bmm-2018-0157","article-title":"Thymidine kinase 1 as a tumor biomarker: technical advances offer new potential to an old biomarker","volume":"12","author":"Jagarlamudi","year":"2018","journal-title":"Biomark Med"},{"key":"ref3","doi-asserted-by":"publisher","first-page":"957","DOI":"10.1292\/jvms.59.957","article-title":"Plasma thymidine kinase activity in dogs with lymphoma and leukemia","volume":"59","author":"Nakamura","year":"1997","journal-title":"J Vet Med Sci"},{"key":"ref4","doi-asserted-by":"publisher","first-page":"1597","DOI":"10.1373\/clinchem.2003.030379","article-title":"Sensitive nonradiometric method for determining thymidine kinase 1 activity","volume":"50","author":"\u00d6hrvik","year":"2004","journal-title":"Clin Chem"},{"key":"ref5","doi-asserted-by":"publisher","first-page":"17","DOI":"10.1016\/j.rvsc.2005.05.001","article-title":"A non-radiometric method for measuring serum thymidine kinase activity in malignant lymphoma in dogs","volume":"80","author":"von Euler","year":"2006","journal-title":"Res Vet Sci"},{"key":"ref6","doi-asserted-by":"publisher","first-page":"40","DOI":"10.1016\/j.tvjl.2012.03.006","article-title":"A sensitive and kinetically defined radiochemical assay for canine and human serum thymidine kinase 1 (TK1) to monitor canine malignant lymphoma","volume":"194","author":"Sharif","year":"2012","journal-title":"Vet J"},{"key":"ref7","doi-asserted-by":"publisher","first-page":"505","DOI":"10.3892\/ijo_00000175","article-title":"Monitoring therapy in canine malignant lymphoma and leukemia with serum thymidine kinase 1 activity\u2014evaluation of a new, fully automated non-radiometric assay","volume":"34","author":"Von Euler","year":"2009","journal-title":"Int J Oncol"},{"key":"ref8","doi-asserted-by":"publisher","first-page":"292","DOI":"10.1111\/j.1476-5829.2011.00298.x","article-title":"Elevated serum thymidine kinase activity in canine splenic hemangiosarcoma","volume":"10","author":"Thamm","year":"2012","journal-title":"Vet Comp Oncol"},{"key":"ref9","doi-asserted-by":"publisher","first-page":"228","DOI":"10.1186\/s12917-014-0228-1","article-title":"Properties of cellular and serum forms of thymidine kinase 1 (TK1) in dogs with acute lymphocytic leukemia (ALL) and canine mammary tumors (CMTs): implications for TK1 as a proliferation biomarker","volume":"10","author":"Jagarlamudi","year":"2014","journal-title":"BMC Vet Res"},{"key":"ref10","doi-asserted-by":"publisher","first-page":"522","DOI":"10.2460\/javma.230.4.522","article-title":"Serum C-reactive protein concentrations in dogs with multicentric lymphoma undergoing chemotherapy","volume":"230","author":"Merlo","year":"2007","journal-title":"J Am Vet Med Assoc"},{"key":"ref11","doi-asserted-by":"publisher","first-page":"28","DOI":"10.1111\/j.1476-5829.2009.00200.x","article-title":"Response evaluation criteria for peripheral nodal lymphoma in dogs (v1.0)\u2014a Veterinary Cooperative Oncology Group (VCOG) consensus document","volume":"8","author":"Vail","year":"2010","journal-title":"Vet Comp Oncol"},{"key":"ref12","doi-asserted-by":"publisher","first-page":"176","DOI":"10.1111\/vco.12032","article-title":"Response evaluation criteria for solid tumours in dogs (v1.0): a Veterinary Cooperative Oncology Group (VCOG) consensus document","volume":"13","author":"Nguyen","year":"2015","journal-title":"Vet Comp Oncol"},{"key":"ref13","doi-asserted-by":"publisher","first-page":"204","DOI":"10.1111\/jvim.16326","article-title":"Performance of lymph node cytopathology in diagnosis and characterization of lymphoma in dogs","volume":"36","author":"Martini","year":"2022","journal-title":"J Vet Intern Med"},{"key":"ref14","doi-asserted-by":"publisher","first-page":"105420","DOI":"10.1016\/j.rvsc.2024.105420","article-title":"Comparison of the accuracy of minimally invasive techniques (cytology, cell block, immunocytochemistry and clonality assay) in the diagnosis of canine multicentric lymphoma","volume":"180","author":"Valente","year":"2024","journal-title":"Res Vet Sci"},{"key":"ref15","doi-asserted-by":"publisher","first-page":"1243853","DOI":"10.3389\/fvets.2023.1243853","article-title":"A monoclonal antibody-based sandwich ELISA for measuring canine thymidine kinase 1 protein and its role as biomarker in canine lymphoma","volume":"10","author":"Sharif","year":"2023","journal-title":"Front Vet Sci"},{"key":"ref16","doi-asserted-by":"publisher","first-page":"373","DOI":"10.1111\/vco.12052","article-title":"Serum thymidine kinase 1 and C-reactive protein as biomarkers for screening clinically healthy dogs for occult disease","volume":"13","author":"Selting","year":"2015","journal-title":"Vet Comp Oncol"},{"key":"ref17","doi-asserted-by":"publisher","first-page":"2565","DOI":"10.3748\/wjg.v25.i21.2565","article-title":"Colorectal cancer screening from 45 years of age: thesis, antithesis and synthesis","volume":"25","author":"Mannucci","year":"2019","journal-title":"World J Gastroenterol"},{"key":"ref18","doi-asserted-by":"publisher","first-page":"e0137871","DOI":"10.1371\/journal.pone.0137871","article-title":"A new sandwich ELISA for quantification of thymidine kinase 1 protein levels in sera from dogs with different malignancies can aid in disease management","volume":"10","author":"Jagarlamudi","year":"2015","journal-title":"PLoS One"},{"key":"ref19","doi-asserted-by":"publisher","first-page":"15785","DOI":"10.1038\/s41598-018-34097-0","article-title":"Multiplexed autoantibody signature for serological detection of canine mammary tumours","volume":"8","author":"Hussain","year":"2018","journal-title":"Sci Rep"},{"key":"ref20","doi-asserted-by":"publisher","first-page":"329","DOI":"10.1186\/s12917-022-03429-8","article-title":"Evaluation of plasma nucleosome concentrations in dogs with a variety of common cancers and in healthy dogs","volume":"18","author":"Wilson-Robles","year":"2022","journal-title":"BMC Vet Res"},{"key":"ref21","doi-asserted-by":"publisher","first-page":"187","DOI":"10.1111\/j.1939-1676.2011.0695.x","article-title":"Mortality in North American dogs from 1984 to 2004: an investigation into age-, size-, and breed-related causes of death","volume":"25","author":"Fleming","year":"2011","journal-title":"J Vet Intern Med"},{"key":"ref22","doi-asserted-by":"publisher","first-page":"535","DOI":"10.3390\/vetsci9100535","article-title":"Vet-OncoNet: malignancy analysis of neoplasms in dogs and cats","volume":"9","author":"Pinello","year":"2022","journal-title":"Vet Sci"},{"key":"ref23","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1080\/01652176.2024.2435978","article-title":"Multiplexed immunoassay for a serum autoantibody biomarker panel in diagnostic and prognostic prediction of canine mammary tumors","volume":"45","author":"Wu","year":"2025","journal-title":"Vet Q"},{"key":"ref24","doi-asserted-by":"publisher","first-page":"101783","DOI":"10.1016\/j.tranon.2023.101783","article-title":"Combination of the biomarkers for aging and cancer?\u2014Challenges and current status","volume":"38","author":"Li","year":"2023","journal-title":"Transl Oncol"},{"key":"ref25","doi-asserted-by":"publisher","first-page":"e0280795","DOI":"10.1371\/journal.pone.0280795","article-title":"Age at cancer diagnosis by breed, weight, sex, and cancer type in a cohort of more than 3,000 dogs: determining the optimal age to initiate cancer screening in canine patients","volume":"18","author":"Rafalko","year":"2023","journal-title":"PLoS One"},{"key":"ref26","doi-asserted-by":"publisher","first-page":"1159","DOI":"10.1111\/jvim.13954","article-title":"Thymidine kinase type 1 and C-reactive protein concentrations in dogs with spontaneously occurring cancer","volume":"30","author":"Selting","year":"2016","journal-title":"J Vet Intern Med"}],"container-title":["Frontiers in Veterinary Science"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fvets.2025.1570106\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,4,25]],"date-time":"2025-04-25T05:26:25Z","timestamp":1745558785000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fvets.2025.1570106\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4,25]]},"references-count":26,"alternative-id":["10.3389\/fvets.2025.1570106"],"URL":"https:\/\/doi.org\/10.3389\/fvets.2025.1570106","relation":{},"ISSN":["2297-1769"],"issn-type":[{"value":"2297-1769","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,4,25]]},"article-number":"1570106"}}