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In this study, a novel Phagomagnetic separation (PhMS)-qPCR assay to rapidly detect viable\n                    <jats:italic>Mycobacterium bovis<\/jats:italic>\n                    in bovine blood samples was evaluated. A total of 149 heart blood samples were collected from cattle at exsanguination point of the slaughter line at a NI abattoir between June and August 2023 \u2013 74 from TB reactor cattle (compulsorily culled within days of a positive SICCT test result) and 75 from routine slaughter cattle (&lt;\u200930 months). Peripheral blood mononuclear cells (PBMCs) were isolated and lysed to release any mycobacteria present before PhMS-qPCR and culture were performed to detect presence of viable\n                    <jats:italic>M. bovis<\/jats:italic>\n                    . The DNAs obtained were subsequently tested for evidence of viable\n                    <jats:italic>Mycobacterium avium<\/jats:italic>\n                    subsp.\n                    <jats:italic>paratuberculosis<\/jats:italic>\n                    (MAP) also. PhMS-qPCR results indicate that of the TB reactor cattle with conclusive PhMS-qPCR results, 7.5% of bloods tested positive for viable\n                    <jats:italic>M. bovis<\/jats:italic>\n                    only, 41.8% for viable MAP only and 28.4% showed evidence of co-infection (both\n                    <jats:italic>M. bovis<\/jats:italic>\n                    and MAP detected). Of the routine slaughter cattle with conclusive PhMS-qPCR results, 22.4% of bloods tested positive for viable\n                    <jats:italic>M. bovis<\/jats:italic>\n                    only, 19.4% for viable MAP only and 20.9% showed evidence of co-infection. Of the blood samples with conclusive culture results, 19.6% TB reactors and 25.4% routine slaughter cattle were\n                    <jats:italic>M. bovis<\/jats:italic>\n                    culture positive. No agreement was observed between the\n                    <jats:italic>M. bovis<\/jats:italic>\n                    PhMS-qPCR assay and\n                    <jats:italic>M. bovis<\/jats:italic>\n                    culture results (Kappa \u2212\u20090.028, 95% CI -0.162 to 0.105). Results of this study provide proof-of-concept that the PhMS-qPCR assay is able to detect viable\n                    <jats:italic>M. bovis<\/jats:italic>\n                    in bovine blood, although a different qPCR assay with greater detection sensitivity will need to be identified going forward. A surprisingly high number of\n                    <jats:italic>M. bovis<\/jats:italic>\n                    \/MAP co-infections were detected in the blood of NI cattle, which may be contributing to failure of the bTB eradication scheme. The latter warrants further investigation.\n                  <\/jats:p>","DOI":"10.1186\/s13620-025-00323-1","type":"journal-article","created":{"date-parts":[[2025,11,29]],"date-time":"2025-11-29T02:17:24Z","timestamp":1764382644000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Evidence of mycobacteriaemias and mycobacterial co-infections uncovered in cattle at slaughter using a novel phage-based PhMS-qPCR assay for viable Mycobacterium bovis and Mycobacterium avium subsp. paratuberculosis"],"prefix":"10.1186","volume":"79","author":[{"given":"Hannah","family":"Dane","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brendan","family":"Gilbride","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Minu","family":"Thomas","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Irene R.","family":"Grant","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,11,29]]},"reference":[{"key":"323_CR1","unstructured":"World Organisation for Animal Health. Chapter 3.4.6. Bovine Tuberculosis. OIE Terrestrial Manual 2018. https:\/\/www.woah.org\/app\/uploads\/2021\/03\/3-04-06-bovine-tb.pdf. Accessed 31 July 2025."},{"key":"323_CR2","doi-asserted-by":"publisher","first-page":"3182","DOI":"10.1017\/S0950268815000291","volume":"143","author":"PA Robinson","year":"2015","unstructured":"Robinson PA. A history of bovine tuberculosis eradication policy in Northern Ireland. Epidemiol Infect. 2015;143:3182\u201395. https:\/\/doi.org\/10.1017\/S0950268815000291.","journal-title":"Epidemiol Infect"},{"key":"323_CR3","unstructured":"TB Partnership Steering Group. Bovine Tuberculosis in Northern Ireland - Blueprint for Eradication. (2025). https:\/\/www.daera-ni.gov.uk\/publications\/tb-partnership-steering-group-bovine-tb-northern-ireland-blueprint-eradication. Accessed 17 July 2025."},{"key":"323_CR4","unstructured":"Department for Agriculture, Environment and Rural Affairs. 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Vet Microbiol. 2011;147(1\u20132):127\u201332. https:\/\/doi.org\/10.1016\/j.vetmic.2010.06.016.","journal-title":"Vet Microbiol"}],"container-title":["Irish Veterinary Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s13620-025-00323-1.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s13620-025-00323-1","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s13620-025-00323-1.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,5]],"date-time":"2026-01-05T16:29:36Z","timestamp":1767630576000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1186\/s13620-025-00323-1"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,29]]},"references-count":42,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,12]]}},"alternative-id":["323"],"URL":"https:\/\/doi.org\/10.1186\/s13620-025-00323-1","relation":{},"ISSN":["2046-0481"],"issn-type":[{"value":"2046-0481","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,11,29]]},"assertion":[{"value":"1 August 2025","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"21 November 2025","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"29 November 2025","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The study protocol was approved by the Faculty of Medicine, Health and Life Sciences Research Ethics Committee, Queen\u2019s University Belfast (REC reference no. MHLS 23_113), in accordance with the review process for Animal Welfare projects.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"Prof Irene Grant is Chief Scientific Officer and co-founder of Rapid-Myco Technologies Limited, a spin-out company from Queen\u2019s University Belfast. The other authors are employees of the spin-out company.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"2"}}