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Applied and Environmental Microbiology, August 1999, p. 3735-3737, Vol. 65, No. 8
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Improved Method for Purification of Bacterial DNA from Bovine Milk for Detection of Brucella spp. by PCR

C. Romero and I. Lopez-Goñi*

Departamento de Microbiología, Universidad de Navarra, 31008 Pamplona, Spain

Received 4 February 1999/Accepted 9 May 1999

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.

* Corresponding author. Mailing address: Departamento de Microbiología, Universidad de Navarra, C/Irunlarrea s/n, 31008 Pamplona, Spain. Phone: (34) 948-425600. Fax: (34) 948-425649. E-mail: ilgoni{at}unav.es.

Applied and Environmental Microbiology, August 1999, p. 3735-3737, Vol. 65, No. 8
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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