{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,29]],"date-time":"2025-09-29T00:09:23Z","timestamp":1759104563690},"reference-count":17,"publisher":"Microbiology Society","issue":"5","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2014,5,1]]},"abstract":"<jats:p>Cholix toxin (ChxA) is an exotoxin reported in <jats:italic>Vibrio cholerae<\/jats:italic> non-O1\/non-O139. Apart from its prototype (ChxA I) we have recently identified two novel variants of this toxin, ChxA II and ChxA III. Our previous investigations indicated that the first two variants may instigate extra-intestinal infections and ChxA II can be more lethal than ChxA I in mice. However, all three cholix toxins (ChxA I to III) failed to show any enterotoxicity in rabbit ileal loops. In this study we developed a PCR-restriction fragment length polymorphism (RFLP) assay to differentiate all three <jats:italic>chxA<\/jats:italic> variants to further understand the importance of each subtype. By using 53 <jats:italic>V. cholerae<\/jats:italic> non-O1\/non-O139 strains harbouring <jats:italic>chxA<\/jats:italic> genes, which were previously categorized by sequencing, and various other strains as negative controls, the PCR-RFLP assay showed 100\u200a% typability and specificity. Furthermore, when applied to differentiate additional <jats:italic>V. cholerae<\/jats:italic> strains, which were also screened for the <jats:italic>chxA<\/jats:italic> gene by colony hybridization, this assay identified <jats:italic>chxA<\/jats:italic> I and <jats:italic>chxA<\/jats:italic> II genes among 18.5\u200a% and 4.5\u200a% of non-O1\/non-O139 strains (<jats:italic>n<\/jats:italic>\u200a=\u200a178), respectively. One non-O1\/non-O139 strain was untypable due to the insertion of an IS<jats:italic>911<\/jats:italic>-like element. Interestingly, the <jats:italic>chxA<\/jats:italic> I gene was detected in 10 out of 137 cholera toxin gene-negative <jats:italic>V. cholerae<\/jats:italic> O1 strains. These results suggest that the PCR-RFLP assay developed in this study can be a rapid and simple method to differentiate the <jats:italic>chxA<\/jats:italic> subtypes.<\/jats:p>","DOI":"10.1099\/jmm.0.070797-0","type":"journal-article","created":{"date-parts":[[2014,2,26]],"date-time":"2014-02-26T03:27:26Z","timestamp":1393385246000},"page":"667-673","source":"Crossref","is-referenced-by-count":6,"title":["Development of a PCR-restriction fragment length polymorphism assay for detection and subtyping of cholix toxin variant genes of Vibrio cholerae"],"prefix":"10.1099","volume":"63","author":[{"given":"Sharda Prasad","family":"Awasthi","sequence":"first","affiliation":[{"name":"International Prevention of Epidemics, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Osaka, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Masahiro","family":"Asakura","sequence":"additional","affiliation":[{"name":"International Prevention of Epidemics, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Osaka, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sucharit Basu","family":"Neogi","sequence":"additional","affiliation":[{"name":"Diarrhoeal Disease Epidemiology and Ecology Department, International Centre for Diarrhoeal Disease Research, Dhaka, Bangladesh"},{"name":"International Prevention of Epidemics, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Osaka, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Atsushi","family":"Hinenoya","sequence":"additional","affiliation":[{"name":"International Prevention of Epidemics, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Osaka, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"T.","family":"Ramamurthy","sequence":"additional","affiliation":[{"name":"National Institute of Cholera and Enteric Diseases, Kolkata, India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shinji","family":"Yamasaki","sequence":"additional","affiliation":[{"name":"International Prevention of Epidemics, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Osaka, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"345","reference":[{"key":"r1","doi-asserted-by":"publisher","DOI":"10.1016\/j.micpath.2007.01.005"},{"key":"r2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.00982-12"},{"key":"r3","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.01317-06"},{"key":"r4","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0409918102"},{"key":"r5","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.88.12.5242"},{"key":"r6","doi-asserted-by":"publisher","DOI":"10.1172\/JCI118114"},{"key":"r7","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.74.5.2937-2946.2006"},{"key":"r8","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-695X.1998.tb01128.x"},{"key":"r9","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M710008200"},{"key":"r10","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6968.2003.tb11522.x"},{"key":"r11","doi-asserted-by":"publisher","DOI":"10.1111\/j.1348-0421.1993.tb03236.x"},{"key":"r12","doi-asserted-by":"publisher","DOI":"10.1111\/j.1472-765X.2010.02895.x"},{"key":"r13","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0510322103"},{"key":"r14","doi-asserted-by":"publisher","DOI":"10.1128\/JB.187.9.2992-3001.2005"},{"key":"r15","doi-asserted-by":"publisher","DOI":"10.1111\/j.1758-2229.2010.00139.x"},{"key":"r16","doi-asserted-by":"publisher","DOI":"10.1017\/S0950268802007392"},{"key":"r17","doi-asserted-by":"publisher","DOI":"10.1111\/j.1574-6968.2007.00957.x"}],"container-title":["Journal of Medical Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.microbiologyresearch.org\/content\/journal\/jmm\/10.1099\/jmm.0.070797-0?crawler=true","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2020,10,6]],"date-time":"2020-10-06T08:17:12Z","timestamp":1601972232000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.microbiologyresearch.org\/content\/journal\/jmm\/10.1099\/jmm.0.070797-0"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2014,5,1]]},"references-count":17,"journal-issue":{"issue":"5","published-print":{"date-parts":[[2014,5,1]]}},"URL":"https:\/\/doi.org\/10.1099\/jmm.0.070797-0","relation":{},"ISSN":["0022-2615","1473-5644"],"issn-type":[{"value":"0022-2615","type":"print"},{"value":"1473-5644","type":"electronic"}],"subject":[],"published":{"date-parts":[[2014,5,1]]}}}