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| Author(s): |
Jaqueline Assumpção Diniz
Total Authors: 1
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| Document type: | Master's Dissertation |
| Press: | São Paulo. |
| Institution: | Universidade de São Paulo (USP). Faculdade de Medicina Veterinária e Zootecnia (FMVZ/SBD) |
| Defense date: | 2018-05-09 |
| Examining board members: |
Lara Borges Keid;
Adriana Cortez;
Amane Paldês Gonçales
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| Advisor: | Lara Borges Keid |
| Abstract | |
Brucella canis is the species that affects dogs, causing reproductive problems and economic losses mainly for the owners of commercial kennels, besides the risk that it represents for the manipulators and the owners of dogs that can acquire the infection by the contact with infected animals or contaminated materials. Several diagnostic methods have been developed for the diagnosis of the infection in dogs. However, the performance of the tests vary regarding sensitivity, specificity, speed and practicality for the identification of infected dogs. Studies indicated that the real-time PCR (rtPCR) might show higher sensitivity and specificity, as well as speed of execution. Therefore, the objective of this study was to develop, standardize and validate a rtPCR assay for the diagnosis of canine brucellosis caused by B. canis using whole blood samples from dogs. A total of 56 dogs from commercial kennels and domiciled animals were tested through blood culturing, serological test, conventional PCR (cPCR) and rtPCR, and subsequently the results obtained in each test were compared using the Kappa coefficient. During the standardization of the rtPCR, three sets of primers and probes (rtPCR-A, B and C) were tested using different annealing temperatures and primer concentrations. rtPCR-B showed better performance under the annealing temperature of 59° C and using 900 nM of primers having detected a higher number of positive dogs when compared to blood culturing and PCRc. However, the test was considered not reliable to be used in canine brucellosis diagnosis, since non-specific reactions occurred in samples from uninfected dogs, suggesting a low specificity of the test. Therefore, the blood culturing and cPCR techniques showed a better performance than the developed rtPCR assay to be used in the diagnosis of B. canis infection in dogs. (AU) | |
| FAPESP's process: | 16/07913-0 - Development, standardization and validation of a Real Time Polymerase Chain Reaction to the diagnosis of canine brucellosis |
| Grantee: | Jaqueline Assumpção Diniz |
| Support Opportunities: | Scholarships in Brazil - Master |