Duplex polymerase chain reaction as a rapid, effective diagnostic test for bovine brucellosis using blood samples
Brucellosis is a major infectious disease of cattle. It is also an international trade barrier for the import and export of dairy and beef products. In Mexico, bovine brucellosis is diagnosed using the card, rivanol, and complement fixation serological tests. Molecular methods such as polymerase chain reaction (PCR) are rapid, specific diagnostic tools for brucellosis. This research developed a duplex PCR assay for the diagnosis of brucellosis in cattle blood samples, using the omp2 and BSCP31 genes. Fifty three (53) blood samples with rivanol titers of 1:400, and 60 serologically-negative samples were used. The optimum concentrations of both primers and magnesium chloride for specific fragment amplifications were 100 nM and 0.5 mM, respectively. The analytical sensitivity of duplex PCR was 100 fg/μ,,,,l DNA, while the optimum amplification concentration was 1 ng/μ,,,,l DNA. Analytical specificity was 100%. Diagnostic sensitivity and specificity were 96.3% and 100%, respectively. The results of this study provide evidence for the routine use of duplex PCR in the diagnosis of bovine brucellosis directly on blood samples, as a highly safe, sensitive, specific method.
| Main Authors: | , , , , , , , , |
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| Format: | Digital revista |
| Language: | spa |
| Published: |
Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias
2012
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| Online Access: | https://cienciaspecuarias.inifap.gob.mx/index.php/Pecuarias/article/view/1814 |
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