Vet World Vol.11 February-2018 Article-18
Research Article
Veterinary World, 11(2): 201-208
https://doi.org/10.14202/vetworld.2018.201-208
Comparison of four polymerase chain reaction assays for the detection of Brucella spp. in clinical samples from dogs
2. Department of Theriogenology, Faculty of Veterinary Science, Chorroarin 280 (1427), Buenos Aires, Argentina.
3. National Health Service and Food Quality (SENASA-DILAB) OIE/Brucellosis Reference Laboratory Talcahuano 1660 (1640), Martinez, Buenos Aires, Argentina.
4. Department of Biotechnology, Center for Research in Veterinary and Agronomic Sciences - National Institute of Agricultural Technology (INTA), N. Repetto y de Los Reseros, 1686, Hurlingham, Buenos Aires, Argentina.
Background and Aim: This study aimed to compare the sensitivity (S), specificity (Sp), and positive likelihood ratios (LR+) of four polymerase chain reaction (PCR) assays for the detection of Brucella spp. in dog's clinical samples.
Materials and Methods: A total of 595 samples of whole blood, urine, and genital fluids were evaluated between October 2014 and November 2016. To compare PCR assays, the gold standard was defined using a combination of different serological and microbiological test. Bacterial isolation from urine and blood cultures was carried out. Serological methods such as rapid slide agglutination test, indirect enzyme-linked immunosorbent assay, agar gel immunodiffusion test, and buffered plate antigen test were performed. Four genes were evaluated: (i) The gene coding for the BCSP31 protein, (ii) the ribosomal gene coding for the 16S-23S intergenic spacer region, (iii) the gene coding for porins omp2a/omp2b, and (iv) the gene coding for the insertion sequence IS711.
Results: The results obtained were as follows: (1) For the primers that amplify the gene coding for the BCSP31 protein: S: 45.64% (confidence interval [CI] 39.81-51.46), Sp: 95.62% (CI 93.13-98.12), and LR+: 10.43 (CI 6.04-18); (2) for the primers that amplify the ribosomal gene of the 16S-23S rDNA intergenic spacer region: S: 69.80% (CI 64.42-75.18), Sp: 95.62 % (CI 93.13-98.12), and LR+: 11.52 (CI 7.31-18.13); (3) for the primers that amplify the omp2a and omp2b genes: S: 39.26% (CI 33.55-44.97), Sp: 97.31% (CI 95.30-99.32), and LR+ 14.58 (CI 7.25-29.29); and (4) for the primers that amplify the insertion sequence IS711: S: 22.82% (CI 17.89 - 27.75), Sp: 99.66% (CI 98.84-100), and LR+ 67.77 (CI 9.47-484.89).
Conclusion: We concluded that the gene coding for the 16S-23S rDNA intergenic spacer region was the one that best detected Brucella spp. in canine clinical samples. Keywords: Brucella, Brucella canis, canine brucellosis, clinical samples, comparison, molecular, polymerase chain reaction.
Keywords: Brucella, Brucella canis, canine brucellosis, clinical samples, comparison, molecular, polymerase chain reaction.
How to cite this article: Boeri EJ, Wanke MM, Madariaga MJ, Teijeiro ML, Elena SA, Trangoni MD (2018) Comparison of four polymerase chain reaction assays for the detection of Brucella spp. in clinical samples from dogs, Veterinary World, 11(2): 201-208.
Received: 12-10-2017 Accepted: 10-01-2018 Published online: 16-02-2018
Corresponding author: Eduardo J. Boeri E-mail: eduardoboeri@hotmail.com
DOI: 10.14202/vetworld.2018.201-208
Copyright: Boeri, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.