A pit-stop semi-nested PCR assay for the detection of toxigenic Vibrio cholerae in environmental water samples was developed and its performance evaluated. The PCR technique amplifies sequences within the cholera toxin operon specific for toxigenic V. cholerae. The PCR procedure coupled with an enrichment culture detected as few as four V. cholerae organisms in pure culture. Treated sewage, surface, ground and drinking water samples were seeded with V, cholerae and following enrichment, a detection limit of as few as 1 V. cholerae cfu ml(-1) was obtained with amplification reactions from crude bacterial lysates. The proposed method, which includes a combination of enrichment, rapid sample preparation and a pit-stop semi-nested PCR, could be applicable in the rapid detection of toxigenic V, cholerae in environmental water samples
Reference:
Theron, J, et al. 2000. Detection of toxigenic vibrio cholera from environmental water samples by an enrichment broth cultivation-pit-stop semi-nested PCR procedure. Journal of Applied Microbiology, vol. 89(3), pp 539-546
Theron, J., Cilliers, J. E., Du Preez, M., Brozel, V., & Venter, S. (2000). Detection of toxigenic vibrio cholera from environmental water samples by an enrichment broth cultivation-pit-stop semi-nested PCR procedure. http://hdl.handle.net/10204/1892
Theron, J, Jacques E Cilliers, M Du Preez, VS Brozel, and SN Venter "Detection of toxigenic vibrio cholera from environmental water samples by an enrichment broth cultivation-pit-stop semi-nested PCR procedure." (2000) http://hdl.handle.net/10204/1892
Theron J, Cilliers JE, Du Preez M, Brozel V, Venter S. Detection of toxigenic vibrio cholera from environmental water samples by an enrichment broth cultivation-pit-stop semi-nested PCR procedure. 2000; http://hdl.handle.net/10204/1892.