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

Authors

  • Fernanda Senter Magajevski FCAV
  • R. J. S. GIRIO Faculdade

DOI:

https://doi.org/10.15361/2175-0106.2008v24n1p29-33

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.

 

Author Biography

Fernanda Senter Magajevski, FCAV

Departamento de Medicina Veterinária Preventiva, FCAV, Unesp

 Lab. de leptospirose e brucelose

Published

30/03/2009

Issue

Section

Microbiology/Microbiologia