{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T14:54:59Z","timestamp":1791903299575,"version":"4.3.4"},"reference-count":35,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T00:00:00Z","timestamp":1791849600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"},{"start":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T00:00:00Z","timestamp":1791849600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Veterinary Medicine &amp;amp; Sci"],"published-print":{"date-parts":[[2026,11]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Rapid, inexpensive and reliable detection of\n                    <jats:italic>Mycobacterium avium<\/jats:italic>\n                    subsp.\n                    <jats:italic>paratuberculosis<\/jats:italic>\n                    (MAP) is essential for the control of Johne's disease. This study developed a rhodamine B hydrazone nanoparticle\u2010based immunosensor coupled with antibody\u2010coated nitrocellulose membranes (IMS\u2010CNM) for the detection of MAP in bovine faecal samples. Anti\u2010MAP antibodies were purified from hyperimmune donkey and rabbit sera. Purified donkey IgG was immobilized on nitrocellulose membranes as the capture antibody, whereas rabbit anti\u2010MAP IgG was conjugated to rhodamine B hydrazone nanoparticles using EDC\u2010NHS chemistry to prepare the chromogenic immunosensor. The diagnostic performance of the assay was evaluated using 89 bovine faecal samples and compared with nested PCR. Nested PCR identified 62 positive and 27 negative samples, whereas the IMS\u2010CNM assay detected 60 positive and 29 negative samples. Using nested PCR as the reference method, the IMS\u2010CNM assay achieved a sensitivity of 90.32% (95% CI: 82.96%\u201397.68%) and a specificity of 85.19% (95% CI: 71.80%\u201398.58%). The analytical limit of detection was approximately 3\u00a0\u00d7\u00a010\n                    <jats:sup>3<\/jats:sup>\n                    MAP cells\/mL in PBS. No significant difference was observed between the IMS\u2010CNM assay and nested PCR in classifying positive and negative samples. The proposed IMS\u2010CNM assay provides a rapid, simple and cost\u2010effective approach for detecting MAP in bovine faeces with diagnostic performance comparable to nested PCR. Following further optimization and validation in larger field studies, this platform may serve as a practical screening tool for Johne's disease, particularly in resource\u2010limited settings.\n                  <\/jats:p>","DOI":"10.1002\/vms3.71278","type":"journal-article","created":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T14:40:41Z","timestamp":1791902441000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Detection of\n                    <i>Mycobacterium avium<\/i>\n                    subsp.\n                    <i>paratuberculosis<\/i>\n                    in Bovine Faeces Using a Rhodamine B Hydrazone Immunosensor on Antibody\u2010Coated Nitrocellulose Membranes"],"prefix":"10.1002","volume":"12","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"}]},{"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"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-4895-2448","authenticated-orcid":false,"given":"Rozhin","family":"Ansari","sequence":"additional","affiliation":[{"name":"Faculty of Veterinary Medicine Shahid Chamran University of Ahvaz  Ahvaz Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9478-260X","authenticated-orcid":false,"given":"Mahdi Pourmahdi","family":"Borujeni","sequence":"additional","affiliation":[{"name":"Department of Food Hygiene Faculty of Veterinary Medicine Shahid Chamran University of Ahvaz  Ahvaz Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"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"}]}],"member":"311","published-online":{"date-parts":[[2026,10,13]]},"reference":[{"key":"e_1_2_12_2_1","doi-asserted-by":"publisher","DOI":"10.22067\/ijvst.2023.83873.1289"},{"key":"e_1_2_12_3_1","doi-asserted-by":"publisher","DOI":"10.31083\/JFSFQ38793"},{"key":"e_1_2_12_4_1","doi-asserted-by":"publisher","DOI":"10.1080\/01652176.2016.1196508"},{"issue":"4","key":"e_1_2_12_5_1","first-page":"302","article-title":"Immunological Response to Mycobacterium avium subsp. paratuberculosis in Chickens","volume":"68","author":"Chui L. 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