Evaluation of Four DNA Extraction Protocols for Brucella abortus Detection by PCR in Tissues from Experimentally Infected Cows with the 2308 Strain

摘要: This study compared 4 protocols for DNA extraction from homogenates of 6 different organs cows infected with the Brucella abortus 2308 strain. The were as follows: GT (guanidine isothiocyanate lysis), Boom (GT lysis carrying suspension diatomaceous earth), PK (proteinase K and Santos (lysis by boiling freezing liquid nitrogen). Positive negative gold standard reference groups generated classical bacteriological methods. All samples processed amplified B4 B5 primers. number positive in placental cotyledons was higher than that other organs. cumulated results showed protocol more sensitive (p=0.003) (p=0.0506) methods similar to method (p=0.2969). resulted false-negative PCR. In conclusion, despite disadvantages methods, best approach direct diagnosis B. includes isolation associated PCR extracted cotyledon or