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Provedor de dados:  BJM
País:  Brazil
Título:  Evaluation of DNA extraction protocols for Brucella abortus pcr detection in aborted fetuses or calves born from cows experimentally infected with strain 2308
Autores:  Matrone,M.
Keid,L.B.
Rocha,V.C.M.
Vejarano,M.P.
Ikuta,C.Y.
Rodriguez,C.A.R.
Ferreira,F.
Dias,R.A.
Ferreira Neto,J.S
Data:  2009-09-01
Ano:  2009
Palavras-chave:  Bovine
Brucellosis
Abortion
PCR
DNA extraction
Resumo:  The objective of the present study was to improve the detection of B. abortus by PCR in organs of aborted fetuses from infected cows, an important mechanism to find infected herds on the eradication phase of the program. So, different DNA extraction protocols were compared, focusing the PCR detection of B. abortus in clinical samples collected from aborted fetuses or calves born from cows challenged with the 2308 B. abortus strain. Therefore, two gold standard groups were built based on classical bacteriology, formed from: 32 lungs (17 positives), 26 spleens (11 positives), 23 livers (8 positives) and 22 bronchial lymph nodes (7 positives). All samples were submitted to three DNA extraction protocols, followed by the same amplification process with the primers B4 and B5. From the accumulated results for organ, the proportion of positives for the lungs was higher than the livers (p=0.04) or bronchial lymph nodes (p=0.004) and equal to the spleens (p=0.18). From the accumulated results for DNA extraction protocol, the proportion of positives for the Boom protocol was bigger than the PK (p<0.0001) and GT (p=0.0004). There was no difference between the PK and GT protocols (p=0.5). Some positive samples from the classical bacteriology were negative to the PCR and viceversa. Therefore, the best strategy for B. abortus detection in the organs of aborted fetuses or calves born from infected cows is the use, in parallel, of isolation by classical bacteriology and the PCR, with the DNA extraction performed by the Boom protocol.
Tipo:  Info:eu-repo/semantics/article
Idioma:  Inglês
Identificador:  http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822009000300010
Editor:  Sociedade Brasileira de Microbiologia
Relação:  10.1590/S1517-83822009000300010
Formato:  text/html
Fonte:  Brazilian Journal of Microbiology v.40 n.3 2009
Direitos:  info:eu-repo/semantics/openAccess
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