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Boeri, Eduardo J.
- Comparison of Four Polymerase Chain Reaction Assays for the Detection of Brucella spp. in Clinical Samples from Dogs
Authors
1 Department of Diagnosis and Biological Products Production, Division of Immunology and Diagnosis, Zoonosis Institute Dr. Luis Pasteur, Av. Diaz Velez 4821 (1405), Buenos Aires, AR
2 Department of Theriogenology, Chorroarín 280 (1427), Buenos Aires, AR
3 National Health Service and Food Quality (SENASA-DILAB) OIE/Brucellosis Reference Laboratory Talcahuano 1660 (1640), Martinez, Buenos Aires, AR
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, AR
Source
Veterinary World, Vol 11, No 2 (2018), Pagination: 201-208Abstract
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.Full Text
