Abstract
Vibrio cholerae O1 and O139 has been known for its ability to cause epidemics. These strains produce cholera toxin which is the main cause of secretory diarrhea. V. cholerae non-O1 and non-O139 strains are also capable of causing gastroenteritis as well as septicemia and peritonitis. It has been proven that virulence factors such as T6SS, hapA, rtxA, and hlyA are present in almost all V. cholerae strains. It is imperative that viable but non-culturable cells of V. cholerae are also detected since they are also known to cause diarrhea. Thus, the aim of this study was to develop an assay that detects all V. cholerae regardless of their serotype, culturable state, and virulence genes present, by targeting the species specific conserved ompW sequence. The developed assay meets these goals with 100% specificity and is capable of detecting as low as 5.46 copy number of V. cholerae. Detection is rapid since neither lengthy incubation period nor electrophoresis is required. The assay had excellent repeatability (CV%: 0.24–1.32) and remarkable reproducibility (CV%: 1.08–3.7). Amplification efficiencies in the 89–100% range were observed. The assay is more economical than Taqman-based multiplex real-time PCR assays. Compared to other real-time assays, the ompW assay is specific and sensitive, has better repeatability and reproducibility, and is more economical.
Highlights
Vibrio cholerae is a Gram-negative, comma shaped facultative pathogen responsible for causing cholera
Vibrio cholerae O1 and O139 serogroups express toxin coregulated pilus which confers the bacteria the ability to colonize the intestine while the cholera toxin is associated with secretory diarrhea [5]
V. cholerae strains were grown in alkaline peptone water for enrichment, and all other strains were grown in nutrient broth for 24 h in 37°C
Summary
Vibrio cholerae is a Gram-negative, comma shaped facultative pathogen responsible for causing cholera. The global incidence of cholera was about 2.8 million cases per year, with 91,000 deaths [1]. V. cholerae O1 has been the etiological agent for several cholera epidemics. The serogroup O139 was responsible for cholera outbreaks in India and other countries in Asia during 1992 [2] and was isolated during the outbreak in November 2000 in India [3] and March–April 2002 in Bangladesh [4]. Vibrio cholerae O1 and O139 serogroups express toxin coregulated pilus which confers the bacteria the ability to colonize the intestine while the cholera toxin is associated with secretory diarrhea [5]. The infectious dose for V. cholerae varies from 106 to 1011 cells [6]
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