{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T11:11:33Z","timestamp":1788952293889,"version":"build-2803163510"},"reference-count":26,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T00:00:00Z","timestamp":1788912000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003624","name":"Ministry of Agriculture, Food and Rural Affairs","doi-asserted-by":"publisher","award":["#RS-2025-02305007"],"award-info":[{"award-number":["#RS-2025-02305007"]}],"id":[{"id":"10.13039\/501100003624","id-type":"DOI","asserted-by":"publisher"}]},{"award":["#RS-2025-02305007"],"award-info":[{"award-number":["#RS-2025-02305007"]}],"id":[{"id":"https:\/\/ror.org\/033w59g46","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Int. J. Mol. Sci.","IJMS"],"abstract":"<jats:p>Canine cancer screening has an important limitation in that organ-specific screening strategies used in human medicine cannot be directly applied across routine veterinary clinical practice. Blood-based volatile organic compound (VOC) profiling may provide a practical approach for pan-cancer screening; however, its application in multicenter and multinational settings requires evaluation that accounts for cohort heterogeneity and analytical variability. In this study, we evaluated the canine cancer screening performance of an integrated classification framework using a headspace gas chromatography\u2013mass spectrometry (HS-GC\/MS)-based targeted blood VOC panel in a retrospective multicohort dataset comprising dogs recruited in Korea and Thailand. Whole-blood samples from 590 dogs recruited in Korea and Thailand were analyzed, including 167 dogs in the malignant tumor group and 423 dogs in the non-cancer control group. The predefined targeted VOC panel consisted of aromatic compounds and straight-chain alkanes and was analyzed using HS-GC\/MS. The Korea and Thailand cohorts were incorporated into a single integrated classification framework rather than being separated into country-specific models. To evaluate performance variability associated with data partitioning, cohort-aware stratified training\u2013validation splitting was repeated 30 times within the integrated retrospective cohort, with both national cohorts represented in the training and validation sets in each repeat. Class imbalance correction was applied only to the training data, and validation performance was assessed using both threshold-dependent and threshold-independent metrics. Across the 30 repeated validations, the integrated framework achieved an AUROC of 0.926 \u00b1 0.019 and an AUPRC of 0.894 \u00b1 0.023. Balanced accuracy, accuracy, sensitivity, specificity, F1-score, MCC, and Cohen\u2019s \u03ba were 0.858 \u00b1 0.024, 0.895 \u00b1 0.018, 0.772 \u00b1 0.048, 0.943 \u00b1 0.022, 0.805 \u00b1 0.033, 0.736 \u00b1 0.046, and 0.734 \u00b1 0.045, respectively. In the sample-level prediction score analysis, the median predicted cancer probabilities for cancer and non-cancer samples were 0.954 and 0.041, respectively, indicating clear probability-based separation between the two groups. In the cohort-specific analysis, the discrimination signal was maintained in both the Korea and Thailand cohorts, although threshold-dependent performance differed between the two cohorts. These findings indicate that an HS-GC\/MS-based targeted blood VOC panel provided a consistent cancer-associated classification signal under repeated internal validation within the specific integrated retrospective dataset evaluated in this study. The results demonstrate the feasibility of evaluating a common classification framework across the Korea and Thailand cohorts without constructing separate country-specific models, while not establishing independent external generalizability or platform-independent robustness. Because the Korea and Thailand cohorts were completely confounded with the Agilent and Thermo Fisher analytical platforms, respectively, the independent contributions of cohort characteristics and analytical platform cannot be determined from the present data. Further independent prospective validation in newly recruited populations and institutions, together with disease-control evaluation including clinically relevant controls, cohort-specific calibration, evaluation across analytical platforms, and assessment of clinical utility within real-world workflows, is required before the external generalizability and clinical applicability of the framework can be determined.<\/jats:p>","DOI":"10.3390\/ijms27188016","type":"journal-article","created":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T10:19:16Z","timestamp":1788949156000},"page":"8016","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["HS-GC\/MS-Based Targeted Blood VOC Panel for Canine Cancer Screening Using an Integrated Classification Framework in Retrospective Korea and Thailand Cohorts"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-4338-2940","authenticated-orcid":false,"given":"Kyung-Geun","family":"Ahn","sequence":"first","affiliation":[{"name":"Center for Veterinary Bioinformatics Research and Development, MetaDx Inc., Hanam 12939, Republic of Korea"},{"name":"Department of Biomedical Laboratory Science, University of Health Science, Eulji University, Uijeongbu 11759, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1556-6480","authenticated-orcid":false,"given":"Jin-Wook Chris","family":"Kim","sequence":"additional","affiliation":[{"name":"MetaDx Inc., Seoul 06248, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Soon Chul","family":"Gwak","sequence":"additional","affiliation":[{"name":"Center for Veterinary Bioinformatics Research and Development, MetaDx Inc., Hanam 12939, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eunkyung","family":"Oh","sequence":"additional","affiliation":[{"name":"Center for Veterinary Bioinformatics Research and Development, MetaDx Inc., Hanam 12939, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seyeon","family":"Lee","sequence":"additional","affiliation":[{"name":"Center for Veterinary Bioinformatics Research and Development, MetaDx Inc., Hanam 12939, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jedsada","family":"Siripoonsub","sequence":"additional","affiliation":[{"name":"Center of Excellence for Companion Animal Cancer, Department of Pathology, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10400, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sirintra","family":"Sirivisoot","sequence":"additional","affiliation":[{"name":"Center of Excellence for Companion Animal Cancer, Department of Pathology, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10400, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patharakrit","family":"Teewasutrakul","sequence":"additional","affiliation":[{"name":"Oncology Clinic, Small Animal Teaching Hospital, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10400, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4888-2677","authenticated-orcid":false,"given":"Somporn","family":"Techangamsuwan","sequence":"additional","affiliation":[{"name":"Department of Pathology, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10400, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Suparat","family":"Chaipipat","sequence":"additional","affiliation":[{"name":"Kasetsart University Veterinary Diagnostic Laboratory\u2014Bangkhen, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kannika","family":"Siripattarapravat","sequence":"additional","affiliation":[{"name":"Kasetsart University Veterinary Diagnostic Laboratory\u2014Bangkhen, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"},{"name":"Department of Pathology, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manakorn","family":"Sukmak","sequence":"additional","affiliation":[{"name":"Department of Farm Resources and Production Medicine, Faculty of Veterinary Medicine, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom 73140, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6137-969X","authenticated-orcid":false,"given":"Anudep","family":"Rungsipipat","sequence":"additional","affiliation":[{"name":"Center of Excellence for Companion Animal Cancer, Department of Pathology, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10400, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giup","family":"Jang","sequence":"additional","affiliation":[{"name":"Center for Veterinary Bioinformatics Research and Development, MetaDx Inc., Hanam 12939, Republic of Korea"},{"name":"MetaDx Inc., Seoul 06248, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5868-1297","authenticated-orcid":false,"given":"Geon A","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Biomedical Laboratory Science, University of Health Science, Eulji University, Uijeongbu 11759, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,9]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"20140231","DOI":"10.1098\/rstb.2014.0231","article-title":"Comparative oncology: What dogs and other species can teach us about humans with cancer","volume":"370","author":"Schiffman","year":"2015","journal-title":"Philos. Trans. R. Soc. Lond. B Biol. Sci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1264","DOI":"10.2105\/AJPH.91.8.1264","article-title":"Colorectal cancer screening participation: Comparisons with mammography and prostate-specific antigen screening","volume":"91","author":"Lemon","year":"2001","journal-title":"Am. J. Public Health"},{"key":"ref_3","first-page":"248","article-title":"Communicating benefits and risks of screening for prostate, colon, and breast cancer","volume":"43","author":"Barrett","year":"2011","journal-title":"Fam. Med."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Ulyte, A., Wei, W., Dressel, H., Gruebner, O., von Wyl, V., B\u00e4hler, C., Blozik, E., Br\u00fcngger, B., and Schwenkglenks, M. (2020). Variation of colorectal, breast and prostate cancer screening activity in Switzerland: Influence of insurance, policy and guidelines. PLoS ONE, 15.","DOI":"10.1371\/journal.pone.0231409"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"103169","DOI":"10.1016\/j.eclinm.2025.103169","article-title":"Congruence of cancer screening recommendations between the USPSTF and the top ten US cancer centers: A cross sectional study","volume":"82","author":"Wright","year":"2025","journal-title":"eClinicalMedicine"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Flory, A., Kruglyak, K.M., Tynan, J.A., McLennan, L.M., Rafalko, J.M., Fiaux, P.C., Hernandez, G.E., Marass, F., Nakashe, P., and Ruiz-Perez, C.A. (2022). Clinical validation of a next-generation sequencing-based multi-cancer early detection \u201cliquid biopsy\u201d blood test in over 1,000 dogs using an independent testing set: The CANcer Detection in Dogs (CANDiD) study. PLoS ONE, 17.","DOI":"10.1371\/journal.pone.0266623"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"258","DOI":"10.1111\/jvim.16616","article-title":"Cancer detection in clinical practice and using blood-based liquid biopsy: A retrospective audit of over 350 dogs","volume":"37","author":"Flory","year":"2023","journal-title":"J. Vet. Intern. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1029","DOI":"10.1126\/science.1160809","article-title":"Understanding the Warburg effect: The metabolic requirements of cell proliferation","volume":"324","author":"Cantley","year":"2009","journal-title":"Science"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Woollam, M., Siegel, A.P., Munshi, A., Liu, S., Tholpady, S., Gardner, T., Li, B.-Y., Yokota, H., and Agarwal, M. (2023). Canine-Inspired Chemometric Analysis of Volatile Organic Compounds in Urine Headspace to Distinguish Prostate Cancer in Mice and Men. Cancers, 15.","DOI":"10.3390\/cancers15041352"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/j.cccn.2004.04.023","article-title":"Diagnostic potential of breath analysis\u2014Focus on volatile organic compounds","volume":"347","author":"Miekisch","year":"2004","journal-title":"Clin. Chim. Acta"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Oyerinde, A.S., Selvaraju, V., Babu, J.R., and Geetha, T. (2023). Potential Role of Oxidative Stress in the Production of Volatile Organic Compounds in Obesity. Antioxidants, 12.","DOI":"10.3390\/antiox12010129"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Mochalski, P., Leja, M., \u015alefarska-Wolak, D., Mezmale, L., Patsko, V., Ager, C., Kr\u00f3licka, A., Mayhew, C.A., Shani, G., and Haick, H. (2023). Identification of Key Volatile Organic Compounds Released by Gastric Tissues as Potential Non-Invasive Biomarkers for Gastric Cancer. Diagnostics, 13.","DOI":"10.3390\/diagnostics13030335"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1186\/s13059-023-03047-z","article-title":"Correcting batch effects in large-scale multiomics studies using a reference-material-based ratio method","volume":"24","author":"Yu","year":"2023","journal-title":"Genome Biol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Ahn, K.-G., Choi, R., Gwak, S., Choi, I., Jang, G., Kim, J.-W., and Kim, G.A. (2025). A Novel Sample Preparation Method for GC-MS Analysis of Volatile Organic Compounds in Whole Blood for Veterinary Use. Int. J. Mol. Sci., 26.","DOI":"10.20944\/preprints202501.1677.v1"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1093\/ckj\/sfaa188","article-title":"External validation of prognostic models: What, why, how, when and where?","volume":"14","author":"Ramspek","year":"2021","journal-title":"Clin. Kidney J."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"e074819","DOI":"10.1136\/bmj-2023-074819","article-title":"Evaluation of clinical prediction models (part 1): From development to external validation","volume":"384","author":"Collins","year":"2024","journal-title":"BMJ"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"771","DOI":"10.1111\/jvim.16075","article-title":"Effect of type of diet on blood and plasma taurine concentrations, cardiac biomarkers, and echocardiograms in 4 dog breeds","volume":"35","author":"Adin","year":"2021","journal-title":"J. Vet. Intern. Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1038\/s41746-025-01543-z","article-title":"A systematic review and meta-analysis of diagnostic performance comparison between generative AI and physicians","volume":"8","author":"Takita","year":"2025","journal-title":"NPJ Digit. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"5007","DOI":"10.1038\/s41467-024-48938-2","article-title":"A unified framework for estimating country-specific cumulative incidence for 18 diseases stratified by polygenic risk","volume":"15","author":"Jermy","year":"2024","journal-title":"Nat. Commun."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1038\/s43856-024-00691-9","article-title":"A unified framework for diagnostic test development and evaluation during outbreaks of emerging infections","volume":"4","author":"Chaturvedi","year":"2024","journal-title":"Commun. Med."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1016\/j.jclinepi.2015.04.005","article-title":"Prediction models need appropriate internal, internal-external, and external validation","volume":"69","author":"Steyerberg","year":"2016","journal-title":"J. Clin. Epidemiol."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Saito, T., and Rehmsmeier, M. (2015). The precision-recall plot is more informative than the ROC plot when evaluating binary classifiers on imbalanced datasets. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0118432"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"981458","DOI":"10.1155\/2015\/981458","article-title":"Current Challenges in Volatile Organic Compounds Analysis as Potential Biomarkers of Cancer","volume":"2015","author":"Schmidt","year":"2015","journal-title":"J. Biomark."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"207","DOI":"10.3109\/00498258409151406","article-title":"Cytochrome P-450 oxidation of alkanes originating as scission products during lipid peroxidation","volume":"14","author":"Remmer","year":"1984","journal-title":"Xenobiotica"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"034001","DOI":"10.1088\/1752-7163\/ab7f9d","article-title":"A mechanistic study and review of volatile products from peroxidation of unsaturated fatty acids: An aid to understanding the origins of volatile organic compounds from the human body","volume":"14","author":"Ratcliffe","year":"2020","journal-title":"J. Breath. Res."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Floss, M.A., Fink, T., Maurer, F., Volk, T., Kreuer, S., and M\u00fcller-Wirtz, L.M. (2022). Exhaled Aldehydes as Biomarkers for Lung Diseases: A Narrative Review. Molecules, 27.","DOI":"10.3390\/molecules27165258"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/18\/8016\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T10:21:10Z","timestamp":1788949270000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/18\/8016"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,9]]},"references-count":26,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["ijms27188016"],"URL":"https:\/\/doi.org\/10.3390\/ijms27188016","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,9]]}}}