{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T02:22:51Z","timestamp":1791944571014,"version":"4.3.4"},"reference-count":55,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2025,3,19]],"date-time":"2025-03-19T00:00:00Z","timestamp":1742342400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005412","name":"Shahid Chamran University of Ahvaz","doi-asserted-by":"publisher","award":["SCU.VP1402.12470"],"award-info":[{"award-number":["SCU.VP1402.12470"]}],"id":[{"id":"10.13039\/501100005412","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Clinical Laboratory Analysis"],"published-print":{"date-parts":[[2025,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Johne's disease, also known as paratuberculosis, is a chronic granulomatous enteritis disease that affects ruminants worldwide.<\/jats:p><\/jats:sec><jats:sec><jats:title>Objective<\/jats:title><jats:p>The objective of this study was to assess the effectiveness of the immunomagnetic bead separation\u2010immunosensor (IMB\u2010IS) detection method compared to Nested\u2010PCR for identifying <jats:styled-content style=\"fixed-case\"><jats:italic>Mycobacterium avium<\/jats:italic><\/jats:styled-content> subspecies <jats:italic>paratuberculosis<\/jats:italic> (MAP) infection in cattle feces samples.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Ninety rectal fecal samples were collected from selected cattle, comprising 59 serum\u2010positive and 31 serum\u2010negative cases based on serum ELISA. Following DNA extraction, nested\u2010PCR was conducted using the IS900 primer sequence targeting the MAP\u2010specific gene. Immunomagnetic bead (IMB) nanoparticles were synthesized by purifying hyperimmune donkey IgG through affinity chromatography and then conjugating it to Fe nanoparticles. Rhodamine\u2010B hydrazone immunosensor (IS) was synthesized and conjugated to hyperimmune rabbit IgG. The synthesized IMB and IS were used to identify MAP in cattle fecal samples.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The results of this study revealed that of the 90 stool samples tested using the nested\u2010PCR method, 62 samples (68.88%) were positive, while 28 samples (31.12%) were negative. In the IMB\u2010IS test based on optical density (OD), 64 samples were positive (71.1%), while 26 samples were negative (28.8%). This test exhibited a sensitivity of 100%, specificity of 92.85%, and an overall test accuracy of 97.77%.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Given the considerations of cost, time, positive and negative predictive values, and acceptable accuracy of the IMB\u2010IS test, it is recommended for evaluation in screening and epidemiological studies.<\/jats:p><\/jats:sec>","DOI":"10.1002\/jcla.70009","type":"journal-article","created":{"date-parts":[[2025,3,19]],"date-time":"2025-03-19T12:24:27Z","timestamp":1742387067000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Comparison of Immunomagnetic Bead Separation\u2010Immunosensor Detection and Nested\u2010<scp>PCR<\/scp> Methods for Detecting <i>Mycobacterium avium<\/i> Subspecies <i>paratuberculosis<\/i> in Cattle Feces"],"prefix":"10.1002","volume":"39","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9510-3464","authenticated-orcid":false,"given":"Mohammad","family":"Khosravi","sequence":"first","affiliation":[{"name":"Department of Pathobiology, Faculty of Veterinary Medicine Shahid Chamran University of Ahvaz  Ahvaz Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9405-2594","authenticated-orcid":false,"given":"Mohammad","family":"Nouri","sequence":"additional","affiliation":[{"name":"Department of Clinical Sciences, Faculty of Veterinary Medicine Shahid Chamran University of Ahvaz  Ahvaz Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8964-8820","authenticated-orcid":false,"given":"Mohammad Rahim","family":"Haji Hajikolaei","sequence":"additional","affiliation":[{"name":"Department of Clinical Sciences, Faculty of Veterinary Medicine Shahid Chamran University of Ahvaz  Ahvaz Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1212-2470","authenticated-orcid":false,"given":"Ali","family":"Kolivand","sequence":"additional","affiliation":[{"name":"Faculty of Veterinary Medicine Shahid Chamran University of Ahvaz  Ahvaz Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3614-0254","authenticated-orcid":false,"given":"Darioush","family":"Gharibi","sequence":"additional","affiliation":[{"name":"Department of Pathobiology, Faculty of Veterinary Medicine Shahid Chamran University of Ahvaz  Ahvaz Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter. 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