{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,31]],"date-time":"2025-12-31T08:05:09Z","timestamp":1767168309077,"version":"build-2238731810"},"reference-count":29,"publisher":"Wiley","license":[{"start":{"date-parts":[[2018,10,2]],"date-time":"2018-10-02T00:00:00Z","timestamp":1538438400000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Canadian Journal of Infectious Diseases and Medical Microbiology"],"published-print":{"date-parts":[[2018,10,2]]},"abstract":"\n Background<\/jats:italic>\n . The conventional methods for diagnosis of\n Vibrio cholerae<\/jats:italic>\n are time consuming, complicated, and expensive. Development of rapid detection tests is critical for prevention and management of cholera. This study aimed to introduce two sensitive sandwich ELISAs based on avian antibodies (IgY) targeting outer membrane protein W (OmpW) and cytotoxin B (CtxB) antigens of\n V. cholerae<\/jats:italic>\n .\n Methods<\/jats:italic>\n . The sequences of\n ompW<\/jats:italic>\n and\n ctxB<\/jats:italic>\n genes were cloned into pET28a vector.\n Escherichia coli<\/jats:italic>\n BL21 (DE3) was transformed with the recombinant vectors, and gene expression was induced by IPTG. The expressed proteins were purified by affinity chromatography using Ni-NTA resins. Two groups of white Leghorn chickens were immunized by recombinant proteins, and the generated antibodies were purified from egg yolks of chickens by PEG precipitation. The antibodies were used for the development of\n \u03b1<\/jats:italic>\n -OmpW and\n \u03b1<\/jats:italic>\n -CtxB ELISAs.\n Results<\/jats:italic>\n . The expression and purification yielded 59 and 38\u2009mg of recombinant OmpW and CtxB, respectively, per one liter of bacterial culture. PEG precipitation and purification of egg yolk antibodies yielded on average (\u00b1SD) 66.5\u2009\u00b1\u20091.80 and 50.9\u2009\u00b1\u20092.23\u2009mg of purified\n \u03b1<\/jats:italic>\n -OmpW and\n \u03b1<\/jats:italic>\n -CtxB per egg, respectively. The analytical sensitivity of\n \u03b1<\/jats:italic>\n -OmpW ELISA was 103\u2009cfu\/mL of\n V. cholerae<\/jats:italic>\n and that of\n \u03b1<\/jats:italic>\n -CtxB ELISA was 33\u2009pg\/mL of recombinant cytotoxin B. The two developed ELISAs did not show any cross-reactivity to any tested bacteria grown in common conditions.\n Discussion<\/jats:italic>\n . The current study is the first report on using IgY for detection of\n V. cholerae<\/jats:italic>\n . The developed ELISAs were shown to have considerable analytical sensitivity and specificity. Therefore, the assays can be one of the convenient methods for sensitive and specific detection of toxigenic\n V. cholerae<\/jats:italic>\n strains in clinical and environmental samples.\n <\/jats:p>","DOI":"10.1155\/2018\/4032531","type":"journal-article","created":{"date-parts":[[2018,10,2]],"date-time":"2018-10-02T19:30:48Z","timestamp":1538508648000},"page":"1-9","source":"Crossref","is-referenced-by-count":8,"title":["Development of IgY-Based Sandwich ELISA as a Robust Tool for Rapid Detection and Discrimination of Toxigenic\n Vibrio cholerae<\/i>"],"prefix":"10.1155","volume":"2018","author":[{"given":"Mahdiye","family":"Bayat","sequence":"first","affiliation":[{"name":"Department of Microbiology, Islamic Azad University-Karaj Branch, Karaj 3148635731, Iran"},{"name":"Diagnostic Biotechnology Unit, Pasteur Institute of Iran, Research and Production Complex, Karaj, Iran"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9461-6118","authenticated-orcid":true,"given":"Alireza","family":"Khabiri","sequence":"additional","affiliation":[{"name":"Diagnostic Biotechnology Unit, Pasteur Institute of Iran, Research and Production Complex, Karaj, Iran"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Behzad","family":"Hemati","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Islamic Azad University-Karaj Branch, Karaj 3148635731, Iran"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","reference":[{"key":"1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.01959-06"},{"issue":"4","key":"2","doi-asserted-by":"crossref","first-page":"1301","DOI":"10.1128\/MMBR.62.4.1301-1314.1998","volume":"62","year":"1998","journal-title":"Microbiology and Molecular Biology 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