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Evaluation of serological, microbiological and molecular markers for the diagnosis of canine brucellosis.

Abstract

Canine brucellosis caused by Brucella canis is chronic infection that affects the reproductive tract of dogs. The infection is important in commercial breeding kennels because once it is introduced in a confined canine population it can reach high prevalence rates, leading to economic losses. Canine brucellosis is a zoonotic infection with public health concern, because of the close contact that is established between dogs and human beings actually. There is no vaccine to be used to prevent canine brucellosis in dogs and the antibiotic treatment of the infection is usually inefficient. Therefore, the laboratory diagnosis comprises an important tool to the control and prevention of the infection because it allows the identification and isolation of the infected animals. The serological tests most commonly used for the diagnosis of canine brucellosis are the rapid slide agglutination test with and without the use of the 2-mercapthoetanol (RSAT and 2ME-RSAT), and the agar gel immunodiffusion test (AGID). However, these serological tests showed a low diagnostic sensitivity and specificity, which difficult the infection diagnosis and compromise the implementation of prophylactic measures. Other serological tests have been used for the diagnosis with variable performances, like the enzyme-linked immunosorbent assay in liquid and in solid phase and the indirect immunofluorescence reaction (IFI). The objective of this project is to standardize the following laboratory tests to the diagnosis of canine brucellosis: an indirect ELISA using the BP26 recombinant antigen from B. abortus, the microbiological culture in lymph node aspirate and conjunctival swab samples, the conventional and real time polymerase chain reaction and loop-mediated isothermal amplification (LAMP) to the direct detection of Brucella DNA in samples of whole blood, lymph node aspirates, semen, vaginal and conjunctival swabs. The results will be compared to the results of the microbiological culture in blood, semen and vaginal swabs and with the results of the serological tests that are commercially available in Brazil. The evaluation will be carried out in naturally infected dogs. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
AGRA BATINGA, MARIA CRYSKELY; RIBEIRO DE LIMA, JULIA TERESA; GREGORI, FABIO; DINIZ, JAQUELINE ASSUMPCAO; MUNER, KERSTIN; OLIVEIRA, TRICIA M. F. S.; FERREIRA, HELENA LAGE; SOARES, RODRIGO MARTINS; KEID, LARA BORGES. Comparative application of IS711-based polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) for canine brucellosis diagnosis. MOLECULAR AND CELLULAR PROBES, v. 39, p. 1-6, JUN 2018. Web of Science Citations: 0.
BATINGA, MARIA CRYSKELY A.; DOS SANTOS, JAINE C.; LIMA, JULIA T. R.; BIGOTTO, MARIA FERNANDO. D.; MUNER, KERSTIN; FAITA, THALITA; SOARES, RODRIGO M.; DA SILVA, DAVID A. V.; OLIVEIRA, TRICIA M. F. S.; FERREIRA, HELENA L.; DINIZ, JAQUELINE A.; KEID, LARA B. Comparison of three methods for recovery of Brucella canis DNA from canine blood samples. Journal of Microbiological Methods, v. 143, p. 26-31, DEC 2017. Web of Science Citations: 2.

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