首页> 外文期刊>Brazilian Journal of Microbiology >Comparison of DNA-extraction methods and Selective Enrichment broths on the detection of Salmonella Typhimurium in swine feces by polymerase chain reaction (PCR)
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Comparison of DNA-extraction methods and Selective Enrichment broths on the detection of Salmonella Typhimurium in swine feces by polymerase chain reaction (PCR)

机译:聚合酶链反应(PCR)检测猪粪中鼠伤寒沙门氏菌的DNA提取方法和选择性富集肉汤比较

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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
机译:这项研究的目的是比较不同的DNA提取方法和选择性富集肉汤通过聚合酶链反应检测人工接种的猪粪样品(100 CFU / g)中鼠伤寒沙门氏菌的有效性。浓缩了Rappaport-Vassiliadis,亚硒酸胱氨酸或Müller-Kauffmann四硫代酸盐后,可通过三种不同的方法提取等分试样:沸腾离心,苯酚-氯仿和盐析。然后将提取的DNA的等分试样用作PCR中的模板。使用沸腾离心和盐析时,选择性富集肉汤对PCR的效率没有影响。另一方面,苯酚-氯仿的效果更好(P

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