Comparison of DNA-extraction methods and Selective Enrichment broths on the detection of Salmonella Typhimurium in swine feces by polymerase chain reaction (PCR)

Brazilian Journal of Microbiology

Carla Roberta Freschi²  Luiz Fernando De Oliveira E Silva Carvalho²  Celso José Bruno De Oliveira¹ 

[1] ,Universidade Estadual Paulista Faculdade de Ciências Agrárias e Veterinárias Departamento de Clínica e Cirurgia VeterináriaJaboticabal SP ,Brasil

关键词: diagnosis;    DNA;    feces;    PCR;    Salmonella Typhimurium;    diagnóstico;    DNA;    fezes;    PCR;    Salmonella Typhimurium;   

DOI  :  10.1590/S1517-83822005000400011

来源: SciELO

【 摘 要 】

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.

【 授权许可】

CC BY-NC   
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