Evaluation of DNA extraction protocols for Brucella abortus pcr detection in aborted fetuses or calves born from cows experimentally infected with strain 2308
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 detectio...
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| Language: | English |
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Sociedade Brasileira de Microbiologia
2009
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| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3768545/ |
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pubmed-37685452013-09-12 Evaluation of DNA extraction protocols for Brucella abortus pcr detection in aborted fetuses or calves born from cows experimentally infected with strain 2308 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 Veterinary Microbiology 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 vice-versa. 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. Sociedade Brasileira de Microbiologia 2009 2009-09-01 /pmc/articles/PMC3768545/ /pubmed/24031391 http://dx.doi.org/10.1590/S1517-838220090003000010 Text en © Sociedade Brasileira de Microbiologia http://creativecommons.org/licenses/by-nc/3.0/ All the content of the journal, except where otherwise noted, is licensed under a Creative Commons License |
| repository_type |
Open Access Journal |
| institution_category |
Foreign Institution |
| institution |
US National Center for Biotechnology Information |
| building |
NCBI PubMed |
| collection |
Online Access |
| language |
English |
| format |
Online |
| author |
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 |
| spellingShingle |
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 Evaluation of DNA extraction protocols for Brucella abortus pcr detection in aborted fetuses or calves born from cows experimentally infected with strain 2308 |
| author_facet |
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 |
| author_sort |
Matrone, M. |
| title |
Evaluation of DNA extraction protocols for Brucella abortus pcr detection in aborted fetuses or calves born from cows experimentally infected with strain 2308 |
| title_short |
Evaluation of DNA extraction protocols for Brucella abortus pcr detection in aborted fetuses or calves born from cows experimentally infected with strain 2308 |
| title_full |
Evaluation of DNA extraction protocols for Brucella abortus pcr detection in aborted fetuses or calves born from cows experimentally infected with strain 2308 |
| title_fullStr |
Evaluation of DNA extraction protocols for Brucella abortus pcr detection in aborted fetuses or calves born from cows experimentally infected with strain 2308 |
| title_full_unstemmed |
Evaluation of DNA extraction protocols for Brucella abortus pcr detection in aborted fetuses or calves born from cows experimentally infected with strain 2308 |
| title_sort |
evaluation of dna extraction protocols for brucella abortus pcr detection in aborted fetuses or calves born from cows experimentally infected with strain 2308 |
| description |
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 vice-versa. 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. |
| publisher |
Sociedade Brasileira de Microbiologia |
| publishDate |
2009 |
| url |
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3768545/ |
| _version_ |
1612010288215752704 |