{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T14:34:50Z","timestamp":1783434890158,"version":"3.54.6"},"reference-count":36,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,1,15]],"date-time":"2025-01-15T00:00:00Z","timestamp":1736899200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81472932"],"award-info":[{"award-number":["81472932"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:p>A colloidal gold immunochromatographic assay (ICA) based on a dual-antibody sandwich method was developed for the rapid and convenient detection of <jats:italic>Orientia tsutsugamushi<\/jats:italic> (<jats:italic>O. tsutsugamushi<\/jats:italic>) antigens in the early stages of infection. Monoclonal antibodies designed as 5B3 targeting the conserved region of 56\u202fkDa outer membrane protein in various strains of <jats:italic>O. tsutsugamushi<\/jats:italic> were generated through cell fusion and screening techniques and combined with previously prepared polyclonal antibodies as detection antibodies to establish the ICA. Colloidal gold and polyclonal antibody-colloidal gold complexes were synthesized under optimized conditions. The nitrocellulose membrane was treated with 5B3 monoclonal antibody and goat anti-mouse antibody as the test and control lines, respectively. The ICA demonstrated robust sensitivity, with a minimum detection limit of 70.5\u202fng for the 56\u202fkDa recombinant of the Gilliam strain. Furthermore, a detection limit of 1\u202f\u00d7\u202f10<jats:sup>6<\/jats:sup> copies\/\u03bcL DNA of <jats:italic>O. tsutsugamushi<\/jats:italic> was determined for both PT and SJ infected cell strains by constructing a relationship between cell number and copy number of the pathogen using a quantitative PCR-based standard curve. The assay also exhibited exceptional specificity, with no false positives observed against other bacterial species, including <jats:italic>Escherichia coli<\/jats:italic>, <jats:italic>Salmonella<\/jats:italic>, <jats:italic>Staphylococcus aureus<\/jats:italic>, and <jats:italic>Listeria monocytogenes<\/jats:italic>. In summary, an ICA which is sensitive, specific, and easy to operate was successfully established for the detection of <jats:italic>O. tsutsugamushi<\/jats:italic> in scrub typhus, potentially enabling early rapid point-of-care diagnosis of scrub typhus.<\/jats:p>","DOI":"10.3389\/fmicb.2024.1521015","type":"journal-article","created":{"date-parts":[[2025,1,15]],"date-time":"2025-01-15T06:12:13Z","timestamp":1736921533000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":5,"title":["Development of a colloidal gold immunochromatographic assay utilizing dual-antibody sandwich method for detecting Orientia tsutsugamushi"],"prefix":"10.3389","volume":"15","author":[{"given":"Qingyu","family":"Lu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shiyin","family":"Yu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sibo","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Min","family":"Cao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liuxin","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miao","family":"Xin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Weilong","family":"Tan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yong","family":"Qi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yichen","family":"Lu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaohui","family":"Xiong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,1,15]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"101888","DOI":"10.1016\/j.cimid.2022.101888","article-title":"Revisiting scrub typhus: a neglected tropical disease","author":"Bhandari","year":"2022","journal-title":"Comp. 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