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The current work highlights the potential application of the developed lateral flow immunochromatographic assay (LFIA) in the rapid and accurate detection of <jats:italic>Brucella<\/jats:italic> spp., underscoring its significance in disease management and control. The study investigated the purity of the extracted <jats:italic>B. abortus<\/jats:italic> smooth lipopolysaccharides-O (S-LPS-O) using high-performance liquid chromatography (HPLC). Moreover, it delved into the development and assessment of a novel diagnostic kit for <jats:italic>Brucella<\/jats:italic> using a recombinant protein A-lateral flow assay in comparison to the RBPT, SAT, MRT, I-ELISA as screening tests, also to C-ELISA, CFT as confirmatory tests. HPLC analysis indicated a distinctive absence of impurities in the extracted S-LPS-O, contrasting with the three peaks observed in the commercial LPS chromatogram. Sensitivity, specificity, and accuracy evaluations were performed, with the LFIA demonstrating promising results, showcasing its potential as a reliable diagnostic tool. Statistical analysis of the LFIA, RBPT, SAT, MRT, ELISA, CFT, and PCR results revealed notable sensitivity, specificity, and accuracy rates, affirming the diagnostic efficacy of the LFIA. The novel layout of the LFIA and the purity of extracted S-LPS-O showcased significant progress in the diagnostic capabilities of the developed LFIA. Moreover, the currently developed LFIA introduces a rapid and reliable diagnostic tool, particularly in resource-limited settings, owing to its ease of use and minimal equipment requirements with its competitively obtained results.<\/jats:p>","DOI":"10.1038\/s41598-025-08741-5","type":"journal-article","created":{"date-parts":[[2025,7,6]],"date-time":"2025-07-06T15:36:44Z","timestamp":1751816204000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Development and evaluation of a novel lateral flow immunoassay for rapid diagnosis of brucellosis across different animal species"],"prefix":"10.1038","volume":"15","author":[{"given":"Mariam","family":"Bedir","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sherif","family":"Marouf","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rafik Hamed","family":"Sayed","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohamed","family":"El-Diasty","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Heba M.","family":"Hassan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Momtaz A.","family":"Shahien","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samah","family":"Eid","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hassan","family":"Aboul-Ella","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jakeen","family":"Eljakee","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,7,6]]},"reference":[{"issue":"5","key":"8741_CR1","doi-asserted-by":"publisher","first-page":"1683","DOI":"10.1128\/aem.62.5.1683-1688.1996","volume":"62","author":"NP Rijpens","year":"1996","unstructured":"Rijpens, N. 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