{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2024,7,31]],"date-time":"2024-07-31T21:40:10Z","timestamp":1722462010996},"reference-count":64,"publisher":"American Society of Tropical Medicine and Hygiene","issue":"6","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2019,6,5]]},"abstract":"<jats:p>The diarrheal disease \u201ccholera\u201d is caused by <jats:italic>Vibrio cholerae<\/jats:italic>, and is primarily confined to endemic regions, mostly in Africa and Asia. It is punctuated by outbreaks and creates severe challenges to public health. The disease-causing strains are most-often members of serogroups O1 and O139. PCR-based methods allow rapid diagnosis of these pathogens, including the identification of their biotypes. However, this necessitates the selection of specific target sequences to differentiate even the closely related biotypes of <jats:italic>V. cholerae<\/jats:italic>. Oligonucleotides for selective amplification of small RNA (sRNA) genes that are specific to these <jats:italic>V. cholerae<\/jats:italic> subtypes were designed. The resulting multiplex PCR assay was validated using <jats:italic>V. cholerae<\/jats:italic> cultures (i.e., 19 <jats:italic>V. cholerae<\/jats:italic> and 22 non\u2013<jats:italic>V. cholerae<\/jats:italic> isolates) and spiked stool samples. The validation using <jats:italic>V. cholerae<\/jats:italic> cultures and spiked stool suspensions revealed detection limits of 10\u2013100 pg DNA per reaction and 1.5 cells\/mL suspension, respectively. The multiplex PCR assay that targets sRNA genes for amplification enables the sensitive and specific detection, as well as the differentiation of <jats:italic>V. cholerae<\/jats:italic>\u2014O1 classical, O1 El Tor, and O139 biotypes. Most importantly, the assay enables fast and cheaper diagnosis compared with classic culture-based methods.<\/jats:p>","DOI":"10.4269\/ajtmh.18-0525","type":"journal-article","created":{"date-parts":[[2019,4,9]],"date-time":"2019-04-09T07:00:51Z","timestamp":1554793251000},"page":"1328-1334","source":"Crossref","is-referenced-by-count":2,"title":["Utilization of Small RNA Genes to Distinguish Vibrio cholerae Biotypes via Multiplex Polymerase Chain Reaction"],"prefix":"10.4269","volume":"100","author":[{"given":"Siti Aminah","family":"Ahmed","sequence":"first","affiliation":[{"name":"Advanced Medical and Dental Institute, Universiti Sains Malaysia, Bertam, Penang, Malaysia;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carsten A.","family":"Raabe","sequence":"additional","affiliation":[{"name":"Institute of Medical Biochemistry, Centre for Molecular Biology of Inflammation (ZMBE), University of M\u00fcnster, M\u00fcnster, Germany;"},{"name":"Brandenburg Medical School (Medizinische Hochschule Brandenburg [MHB]), Neuruppin, Germany;"},{"name":"Institute of Experimental Pathology (ZMBE), University of M\u00fcnster, M\u00fcnster, Germany;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hong Leong","family":"Cheah","sequence":"additional","affiliation":[{"name":"Advanced Medical and Dental Institute, Universiti Sains Malaysia, Bertam, Penang, Malaysia;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chee Hock","family":"Hoe","sequence":"additional","affiliation":[{"name":"Advanced Medical and Dental Institute, Universiti Sains Malaysia, Bertam, Penang, Malaysia;"},{"name":"Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, Kelantan, Malaysia;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Timofey S.","family":"Rozhdestvensky","sequence":"additional","affiliation":[{"name":"Medical Faculty, Transgenic Animal and Genetic Engineering Models (TRAM), University of M\u00fcnster, M\u00fcnster, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thean Hock","family":"Tang","sequence":"additional","affiliation":[{"name":"Advanced Medical and Dental Institute, Universiti Sains Malaysia, Bertam, Penang, Malaysia;"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"2795","reference":[{"key":"b1","series-title":"PLoS Negl Trop Dis","first-page":"e0003832","article-title":"Updated global burden of cholera in endemic countries","volume":"9","author":"Ali","year":"2015"},{"key":"b2","series-title":"Wkly Epidemiol Rec","first-page":"433","article-title":"Cholera annual report 2015","volume":"91","author":"World Health Organization","year":"2016"},{"key":"b3","series-title":"Intern J Sci Res","first-page":"314","article-title":"Using fuzzy classifier for cholera analysis","volume":"3","author":"Okpor","year":"2012"},{"key":"b4","series-title":"Nat Rev Microbiol","first-page":"693","article-title":"Cholera transmission: the host, pathogen and 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