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case_images: (A) PCR analysis for toxR gene of genomic DNA of V. cholerae non-O1/O139 strain isolated from urine specimen. Lane M, 100 base pair (bp) DNA ladder; lane 1, V. cholerae O1 Ogawa-Classical NIH41 (positive control for toxR gene); lane 2, negative control; lane 3, V. cholerae non-O1/O139 strain isolated from urine specimen. (B) PCR analysis for ompU gene of genomic DNA of V. cholerae non-O1/O139 strain isolated from urine specimen. Lane M, 100 base pair (bp) DNA ladder; lane 1, V. cholerae O1 Ogawa-Classical NIH41 (positive control for ompU gene); lane 2, negative control; lane 3, V. cholerae non-O1/O139 strain isolated from urine specimen.
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case_images: France), and the isolate was resistant to ampicillin and susceptible to ciprofloxacin and trimethoprim-sulfamethoxazole. The isolate was further confirmed as V. cholerae by PCR targeting the V. cholerae species specific toxR (Figure 1A) and ompU genes (Figure 1B) and strain tested positive for both genes. Furthermore, the strain was tested positive for major cholera toxin (ctx) gene and lack the TCP gene which is required for colonization (Figure 2).
case_images: Multiplex-PCR analysis for ctxA and tcpA genes. Lane M, 100 bp DNA ladder; lane 1, V. cholerae O1 Ogawa-Classical NIH41 (positive control for ctxA and tcpA gene); lane 2, negative control; lane 3, V. cholerae non-O1/O139 strain isolated from urine specimen.
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case_images: France), and the isolate was resistant to ampicillin and susceptible to ciprofloxacin and trimethoprim-sulfamethoxazole. The isolate was further confirmed as V. cholerae by PCR targeting the V. cholerae species specific toxR (Figure 1A) and ompU genes (Figure 1B) and strain tested positive for both genes. Furthermore, the strain was tested positive for major cholera toxin (ctx) gene and lack the TCP gene which is required for colonization (Figure 2).
case_images: Multiplex-PCR analysis for zot, ace and hlyA. Lane M, 100 bp DNA ladder; lane 1, V. cholerae O1 Ogawa-Classical NIH41 (positive control for ctxA and tcpA gene); lane 2, negative control; lane 3, V. cholerae non-O1/O139 strain isolated from urine specimen.
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case_images: Multiplex-PCR analysis revealed V. cholerae strain was harboring gene encoding hemolysin (hlyA) which play significant role in disease mechanism and invasion, while it was lacking the zonular occludens toxin (zot) and accessory colonization enterotoxin (ace) genes (Figure 3).
case_images: Dendrogram is based on the unweighted-pair group method with arithmetic means (UPGMA) and Pearson correlation coefficient, showing genetic similarity among toxigenic V. cholerae non-O1/O139 isolated from urine and reference control strains of V. cholerae O1 Ogawa-Classical NIH41, V. cholerae O1 Inaba-Classical NIH35A3, and V. cholerae O139 ATCC 5139. All strains were typed by ERIC-PCR DNA fingerprinting and electrophoretic patterns analyzed (UPGMA and Pearson correlation coefficient) with the Gelj software. Two control strains of V. cholerae O1 Ogawa-Classical NIH41 and V. cholerae O1 Inaba-Classical NIH35A3 with 100% genetic similarity, whereas toxigenic strain of V. cholerae non-O1/O139 isolated from urine showed 90% genetic similarity homogenous with control strain of V. cholerae O139 ATCC 5139.
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case_images: ERIC-PCR was used as genomic fingerprinting for determination of clonality and relatedness of isolated V. cholerae strain from urine with three V. cholerae control strains (Figure 4). Based on analyzed result of ERIC fingerprint patterns using UPGMA and Pearson correlation coefficient, isolated V. cholerae strains share 90% genetic similarity with the control strain of V. cholerae O139 ATCC 5139 (Figure 4).