Improved method for purification of bacterial DNA from bovine milk for detection of Brucella spp. by PCR.
Different methods of extraction of bacterial DNA from bovine milk to improve the direct detection of Brucella by PCR were evaluated. We found that the use of a lysis buffer with high concentrations of Tris, EDTA, and NaCl, high concentrations of sodium dodecyl sulfate and proteinase K, and high temperatures of incubation was necessary for the efficient extraction of Brucella DNA. The limit of detection by PCR was 5 to 50 Brucella CFU/ml of milk.
Romero
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C
Departamento de Microbiología, Universidad de Navarra, 31008 Pamplona, Spain.
Lopez-Goñi
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I
D000818:Animals; D001431:Bacteriological Techniques; D002002:Brucella / Q000235:genetics* / Q000302:isolation & purification*; D002417:Cattle; D015169:Colony Count, Microbial; D004269:DNA, Bacterial / Q000235:genetics* / Q000302:isolation & purification*; D005260:Female; D008892:Milk / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods
D004269:DNA, Bacterial