Effect of different enrichment media and DNA extraction techniques on Salmonella detection by PCR in SWINE feces
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The SafePork conference series began in 1996 to bring together international researchers, industry, and government agencies to discuss current Salmonella research and identify research needs pertaining to both pig and pork production. In subsequent years topics of research presented at these conferences expanded to include other chemical and biological hazards to pig and pork production.
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Abstract
The aim of this study was to evaluate different selective enrichment broths and DNA extraction techniques on the detection of Salmonella Typhimurium in swine feces by PCR. Feces samples (n=10) were inoculated with approximately 102 Salmonella Typhimurium organisms, first enriched in GN-Hajna broth and secondly enriched in Rappaport-Vassiliadis, Muller-Kaufmann tetrathionate and selenite-cystine broths. In order to produce DNA-templates for PCR, aliquots from the broths were subjected to three DNA extraction methods: boiling-centrifugation, salting-out and phenol-chloroform. Detection of Salmonella was significant lower when phenol-chloroform was applied to selenite-cystine and Muller-Kaufmann tetrathionate (P<0.05). The boiling-centrifugation technique had best cost/benefit ratio and can be successfully used as a rapid DNA template preparation from the three enriched broths tested.