Comparison of Dna-extraction Methods and Selective Enrichment Broths on the Detection of Salmonella Typhimurium in Swine Feces by Polymerase Chain Reaction (pcr)

نویسندگان

  • Carla Roberta Freschi
  • Celso José
  • Bruno de Oliveira
چکیده

The aim of this study was to compare different DNA-extraction methods and selective enrichment broths for their effectiveness to detect Salmonella Typhimurium in artificially inoculated swine feces samples (100 CFU/ g) by polymerase chain reaction. After enrichment in Rappaport-Vassiliadis, selenite cystine or MüllerKauffmann tetrathionate, aliquots were used for DNA extraction by three different methods: boilingcentrifugation, phenol-chloroform and salting-out. Aliquots of extracted DNA were then used as template in PCR. The selective enrichment broths had no effect on the efficiency of PCR when boiling-centrifugation and salting-out were used. On the other hand, phenol-chloroform was superior (P<0.05) when combined to Rappaport-Vassiliadis. Considering cost and efficiency parameters, we encourage the use of Müller-Kauffmann tetrathionate broth in combination with boiling-centrifugation DNA-extraction procedure.

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تاریخ انتشار 2006