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Brazilian Journal of Microbiology

versão impressa ISSN 1517-8382versão On-line ISSN 1678-4405

Resumo

FRESCHI, Carla Roberta; CARVALHO, Luiz Fernando de Oliveira e Silva  e  OLIVEIRA, Celso José Bruno de. Comparison of DNA-extraction methods and Selective Enrichment broths on the detection of Salmonella Typhimurium in swine feces by polymerase chain reaction (PCR). Braz. J. Microbiol. [online]. 2005, vol.36, n.4, pp.363-367. ISSN 1517-8382.  http://dx.doi.org/10.1590/S1517-83822005000400011.

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 Müller-Kauffmann tetrathionate, aliquots were used for DNA extraction by three different methods: boiling-centrifugation, 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.

Palavras-chave : diagnosis; DNA; feces; PCR; Salmonella Typhimurium.

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