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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Comparison of three methods for recovery of Brucella canis DNA from canine blood samples

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Author(s):
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Batinga, Maria Cryskely A. [1] ; dos Santos, Jaine C. [2] ; Lima, Julia T. R. [1] ; Bigotto, Maria Fernando. D. [2] ; Muner, Kerstin [2] ; Faita, Thalita [2] ; Soares, Rodrigo M. [1] ; da Silva, David A. V. [1] ; Oliveira, Tricia M. F. S. [2] ; Ferreira, Helena L. [2] ; Diniz, Jaqueline A. [1] ; Keid, Lara B. [2]
Total Authors: 12
Affiliation:
[1] Univ Sao Paulo, Fac Med Vet & Zootecnia, Dept Med Vet Prevent & Saude Anim, Pirassununga - Brazil
[2] Univ Sao Paulo, Fac Zootecnia & Engn Alimentos, Dept Med Vet, Pirassununga, SP - Brazil
Total Affiliations: 2
Document type: Journal article
Source: Journal of Microbiological Methods; v. 143, p. 26-31, DEC 2017.
Web of Science Citations: 2
Abstract

Brucella canis, a gram-negative, facultative intracellular and zoonotic bacterium causes canine brucellosis. Direct methods are the most appropriate for the detection of canine brucellosis and bacterial isolation from blood samples has been employed as gold-standard method. However, due to the delay in obtaining results and the biological risk of the bacterial culturing, the polymerase chain reaction (PCR) has been successfully used as an alternative method for the diagnosis of the infection. Sample preparation is a key step for successful PCR and protocols that provide high DNA yield and purity are recommended to ensure high diagnostic sensitivity. The objective of this study was to evaluate the performance of PCR for the diagnosis of B. canis infection in 36 dogs by testing DNA of whole blood obtained through different extraction and purification protocols. Methods 1 and 2 were based on a commercial kit, using protocols recommended for DNA purification of whole blood and tissue samples, respectively. Method 3 was an in-house method based on enzymatic lysis and purification using organic solvents. The results of the PCR on samples obtained through three different DNA extraction protocols were compared to the blood culture. Of the 36 dogs, 13 (36.1%) were positive by blood culturing, while nine (25.0%), 14 (38.8%), and 15 (41.6%) were positive by PCR after DNA extraction using methods 1, 2 and 3, respectively. PCR performed on DNA purified by Method 2 was as efficient as blood culturing and PCR performed on DNA purified with in-house method, but had the advantage of being less laborious and, therefore, a suitable alternative for the direct B. canis detection in dogs. (AU)

FAPESP's process: 15/06072-9 - Evaluation of serological, microbiological and molecular markers for the diagnosis of canine brucellosis.
Grantee:Lara Borges Keid
Support Opportunities: Regular Research Grants