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However, molecular assays such as colourimetric loop-mediated isothermal amplification (LAMP) are often limited by laboratory-based DNA extraction. This study evaluated a simplified HotSHOT (HS) DNA preparation method for extracting\n                    <jats:italic>E. coli<\/jats:italic>\n                    DNA from bovine milk and assessed its compatibility with colourimetric LAMP and conventional PCR. Pasteurised bovine milk spiked with\n                    <jats:italic>E. coli<\/jats:italic>\n                    was used for analytical evaluation, and culture\u2011positive clinical milk samples were tested to assess diagnostic performance. The HS method was compared with five commercial DNA extraction kits and a direct boiling method without centrifugation. Performance was assessed using limit of detection (LOD) expressed as colony-forming units per millilitre. Using the HS method, the LOD was 7.9\u2009\u00d7\u200910\u2074 CFU\/mL for LAMP and 7.9\u2009\u00d7\u200910\u00b3 CFU\/mL for PCR. PCR performance of the HS method was comparable to the QIAamp and Norgen commercial kits, while LAMP sensitivity was ten- to a hundred-fold lower than the best kit-based methods. Direct boiling showed markedly reduced sensitivity, with an LOD of 7.9\u2009\u00d7\u200910\u2078 CFU\/mL. Among 17 culture-confirmed clinical mastitis milk samples positive for\n                    <jats:italic>E. coli<\/jats:italic>\n                    , LAMP detected 15 (88.2%) and PCR detected all 17 (100%). These results show that simplified DNA preparation can support molecular detection of\n                    <jats:italic>E. coli<\/jats:italic>\n                    in bovine milk and provide a practical basis for future development of field-deployable diagnostics in resource-constrained settings.\n                  <\/jats:p>","DOI":"10.1007\/s11259-026-11576-8","type":"journal-article","created":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T05:02:50Z","timestamp":1791954170000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Colourimetric LAMP detection of Escherichia coli in bovine milk using a simplified DNA preparation method"],"prefix":"10.1007","volume":"51","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2955-5638","authenticated-orcid":false,"given":"Tewodros Fentahun","family":"Jember","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7400-1960","authenticated-orcid":false,"given":"Mark Edward","family":"Westman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0672-4478","authenticated-orcid":false,"given":"Sameer Dinkar","family":"Pant","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9820-5308","authenticated-orcid":false,"given":"Seyed Ali","family":"Ghorashi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,10,14]]},"reference":[{"key":"11576_CR1","doi-asserted-by":"crossref","DOI":"10.3389\/fvets.2022.800322","volume":"9","author":"DJ Ajose","year":"2022","unstructured":"Ajose DJ, Oluwarinde BO, Abolarinwa TO, Fri J, Montso KP, Fayemi OE, Aremu AO, Ateba CN (2022) Combating bovine mastitis in the dairy sector in an era of antimicrobial resistance: ethno-veterinary medicinal option as a viable alternative approach. 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