TOMA DE MUESTRAS FRENTE A MORTANDADES DE PECES
Porcine brucellosis is an important zoonosis to public health and to agricultural economy since it causes significant losses in production, and the establishment of sanitary restrictions on local and international trade. Therefore, it is important to have a rapid and reliable diagnosis. To date, con...
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| Formato: | Tesis de maestría acceptedVersion |
| Lenguaje: | Español |
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Universidad de Buenos Aires. Facultad de Ciencias Veterinarias
2015
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| Acceso en línea: | http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=avemaster&cl=CL1&d=HWA_1274 https://repositoriouba.sisbi.uba.ar/gsdl/collect/avemaster/index/assoc/HWA_1274.dir/1274.PDF |
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| Sumario: | Porcine brucellosis is an important zoonosis to public health and to agricultural economy since it causes significant losses in production, and the establishment of sanitary restrictions on local and international trade. Therefore, it is important to have a rapid and reliable diagnosis. To date, confirmatory diagnosis of brucellosis mainly relies on the isolation of the bacterium, but it is difficult to implement. Currently, most of the diagnostic tests are based on serological assays useful for diagnosing herds; however there is still not an accurate diagnosis to test individual animals. Thus, in the present work we developed and evaluated a novel indirect immunoassay (iELISA) using a recombinant glycoprotein antigen formed by the LPS O: 9-O polysaccharide of Yersinia enterocolitica (identical to the O polysaccharide of B. suis) covalently coupled to the carrier protein AcrA, derived from Campylobacter jejuni. This antigen was previously characterized and validated for the diagnosis of brucellosis in humans and cattle. From the above mentioned antigen we have developed and validated the glyco-iELISA using a panel of positive and negative sera previously characterized by agglutination buffered plate antigen techniques (BPA), and by fluorescent polarization assay (FPA). The results indicate that the glyco-iELISA differentiates positive animals from clearly negative ones. A cut-off value of 0.56 resulted in a diagnostic test sensitivity and specificity of 100 %, respectively. In addition, as a proof of concept we set to develop, with a small panel of sera, an indirect fluorescence immunoassay with OAg-AcrA coupled to magnetic beads. Due to the advantage of using magnetic racks instead of centrifugation, reduced incubation time and more efficient washes, this platform has shown to be a valuable practical diagnostic tool. Both developed assays discriminate positive from negative animals, even in infected herds specifically identifying the infected animal and differentiating it from negative animals exposed.
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