Facile and Rapid Detection of Vibrio cholerae by Multiplex PCR Based on ompU, ctxA, and toxR Genes

Several epidemic and endemic cases have been reported involving Vibrio cholerae (V. cholerae) as a causative organism in serious diarrheal diseases with high mortality. Hence, quick identification of this microorganism is critical for health care communities.. This investigation suggests an accurate, sensitive and rapid test to detect toxigenic and pathogenic species of V. cholerae simultaneously.. The standard bacteriological tests were utilized to isolate V. cholerae from the clinical samples. A multiplex PCR assay was carried out with three separate primers designed for ctxA, toxR and ompU genes, which play basic roles in toxigenicity and pathogenicity. To investigate the specificity of the designed primers, the amplification statuses of Salmonella typhi (S. typhi), Shigella dysenteriae (S. dysenteriae) and Escherichia coli O157:H7, as common intestinal pathogens, were evaluated. In addition, different concentrations of V. cholerae genome were used for determining the test sensitivity.. From the total of collected stool samples, 72 V. cholerae isolates were found and separated from the patients. All of them carried toxR and ompU genes, but ctxA gene was detected in only 61 isolates. The specificity and sensitivity of this test was 100%.. Obtained data demonstrated that the designed assay is an accurate, easy, rapid, and cost-effective tool for detecting V. cholerae that can serve as an alternate to current bacteriological tests..