EVALUATION OF THE SENSIBILITY OF PCR USING FOUR METHODS OF DNA EXTRACTION FROM BULL SEMEN EXPERIMENTALLY CONTAMINATED WITH Leptospira interrogans SEROVAR POMONA.

Fernanda Senter Magajevski, R. J. S. GIRIO

Abstract



 


 


Because of the great concern about the sanitary quality of semen in artificial insemination centers all around the world, this study was aimed at evaluating the sensibility of polimerase chain reaction (PCR) using four methods for extraction of DNA in bovine semen experimentally contaminated with Leptospira interrogans serovar Pomona. The primers were Lep 1 and Lep 2, and the techniques of extraction were enzymatic lysis with proteinase K, diatomaceous earth, boiling phenol lysis, and DNAzol. The first method was more effective and showed the best results when semen was diluted 1:3 in TE. These results evidence that choosing the correct DNA extraction procedure is essential to success in the search for etiological agents by PCR.


KEY-WORDS: Leptospirosis. Bovine. Semen. PCR.




 





DOI: http://dx.doi.org/10.15361/2175-0106.2008v24n1p29-33