Theron, JMorar, DDu Preez, MBrozel, VSVenter, SN2007-02-062007-06-072007-02-062007-06-072001-03Theron, J, et al. 2001. Sensitive seminested PCR method for the detection of Shigella in spiked environmental water samples. Water Research, vol. 35(4), pp 869-8740043-1354http://hdl.handle.net/10204/1505http://hdl.handle.net/10204/1505A rapid seminested polymerase chain reaction (PCR) method for the specific, sensitive detection of virulent Shigella spp. in spiked environmental water samples was developed. A set of primers specific for the invasion plasmid antigen gene (ipaH) of virulent Shigella spp, and enteroinvasive Escherichia coli produced a 620-bp fragment that was used as template for the seminested primer pair delineating a 401-bp fragment. By using agarose gel electrophoresis for detection of the seminested PCR-amplified products, a detection limit of 1.6 X 10(3) cfu S. flexneri was obtained with amplification reactions from crude bacterial lysates. The PCR procedure coupled with an enrichment culture incubated for 6 h detected as few as 1.6 S. flexneri organisms in pure culture. Treated sewage, ground, surface and drinking water samples collected from various sources were seeded with S. flexneri and incubated in GN broth for 6 h before detection by seminested PCR. A detection limit lower than 14 cfu/ml was achieved in some water samples. The results indicate that the described seminested PCR has the advantage of a rapid turnaround time and it fulfills the requirements of sensitivity and specificity for use in an environmental laboratory.166599 bytesapplication/pdfenCopyright: 2001 Pergamon-Elsevier Science LtdShigellaSeminested PCRPolymerase chain reactionEnteroinvasive EEnvironmental water samplesEnvironmental sciencesWater resourcesArticleTheron, J., Morar, D., Du Preez, M., Brozel, V., & Venter, S. (2001). Sensitive seminested PCR method for the detection of Shigella in spiked environmental water samples. http://hdl.handle.net/10204/1505Theron, J, D Morar, M Du Preez, VS Brozel, and SN Venter "Sensitive seminested PCR method for the detection of Shigella in spiked environmental water samples." (2001) http://hdl.handle.net/10204/1505Theron J, Morar D, Du Preez M, Brozel V, Venter S. Sensitive seminested PCR method for the detection of Shigella in spiked environmental water samples. 2001; http://hdl.handle.net/10204/1505.TY - Article AU - Theron, J AU - Morar, D AU - Du Preez, M AU - Brozel, VS AU - Venter, SN AB - A rapid seminested polymerase chain reaction (PCR) method for the specific, sensitive detection of virulent Shigella spp. in spiked environmental water samples was developed. A set of primers specific for the invasion plasmid antigen gene (ipaH) of virulent Shigella spp, and enteroinvasive Escherichia coli produced a 620-bp fragment that was used as template for the seminested primer pair delineating a 401-bp fragment. By using agarose gel electrophoresis for detection of the seminested PCR-amplified products, a detection limit of 1.6 X 10(3) cfu S. flexneri was obtained with amplification reactions from crude bacterial lysates. The PCR procedure coupled with an enrichment culture incubated for 6 h detected as few as 1.6 S. flexneri organisms in pure culture. Treated sewage, ground, surface and drinking water samples collected from various sources were seeded with S. flexneri and incubated in GN broth for 6 h before detection by seminested PCR. A detection limit lower than 14 cfu/ml was achieved in some water samples. The results indicate that the described seminested PCR has the advantage of a rapid turnaround time and it fulfills the requirements of sensitivity and specificity for use in an environmental laboratory. DA - 2001-03 DB - ResearchSpace DP - CSIR KW - Shigella KW - Seminested PCR KW - Polymerase chain reaction KW - Enteroinvasive E KW - Environmental water samples KW - Environmental sciences KW - Water resources LK - https://researchspace.csir.co.za PY - 2001 SM - 0043-1354 T1 - Sensitive seminested PCR method for the detection of Shigella in spiked environmental water samples TI - Sensitive seminested PCR method for the detection of Shigella in spiked environmental water samples UR - http://hdl.handle.net/10204/1505 ER -