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Madariaga, Maria 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.