Comparison of four polymerase chain reaction assays for the detection of Brucella spp. in clinical samples from dogs

Boeri, Eduardo; Wanke, M.M.; Madariaga, María Julia; Teijeiro, María Luisa; Elena, Sebastián; Trangoni, Marcos David

doi:10.14202/vetworld.2018.201-208

Cited by 15 publications

(10 citation statements)

References 26 publications

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“…Bacterial isolation has been considered the only proof of B. canis infection (39, 60, 62). More recently, PCR of clinical samples has been suggested as a diagnostic test for detection of subclinical and early infections prior to antibody development (63–67). PCR detects Brucella spp.…”

Section: Discussionmentioning

confidence: 99%

Canine Brucellosis: Insights Into the Epidemiologic Situation in Europe

Buhmann¹,

Paul

Herbst

et al. 2019

Front. Vet. Sci.

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Brucella canis is one of many responsible pathogens of discospondylitis in dogs and infections require specific management. Little is known about the epidemiologic situation in Europe. The purpose of the study was to get insights into the occurrence of brucellosis in dogs in Europe. The database of a European veterinary laboratory was screened for Brucella positive samples. Additionally, medical records of a veterinary hospital in Germany were screened for diagnosis of discospondylitis and brucellosis. The laboratory received samples from 20 European countries for Brucella testing in dogs: 3.7% of submitted samples were Brucella spp. PCR-positive (61/1,657), and Brucella canis antibodies were identified in 5.4% of submitted samples (150/2,764). Brucella spp. PCR-positive samples originated from Spain (11.1% of submitted samples), Poland (6.7% of submitted samples) and rarely from Italy and France. Samples with Brucella canis antibodies originated from 13 European countries (Sweden, Belgium, Austria, Switzerland, Italy, Finland, Germany, Denmark, Hungary, Norway, Poland, France, Netherlands). Young dogs (0–24 months) had a 5.4-fold increased risk of PCR positive samples. The supplementary medical records search identified four young female dogs (7–30 months) with Brucella canis discospondylitis in Germany. The four dogs had been imported to Germany from Eastern European countries (Moldavia, Romania, Macedonia). In conclusion, infection with Brucella canis needs to be considered in dogs in Europe and diagnostics for Brucella canis infection appear indicated in young dogs with discospondylitis.

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Section: Discussionmentioning

confidence: 99%

Canine Brucellosis: Insights Into the Epidemiologic Situation in Europe

Buhmann¹,

Paul

Herbst

et al. 2019

Front. Vet. Sci.

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“…Researchers have paid close attention to diagnostic methods based on qPCR because they are rapid, specific, and sensitive. Some species of Brucella are not harmful to human health, such as Brucella canis and Brucella suis, but veterinarians have developed PCR-based assays for their detection (Boeri et al, 2018;Sabrina et al, 2018). However, the course of brucellosis and the endpoint of the treatment are not the primary issues to veterinarians.…”

Section: Discussionmentioning

confidence: 99%

Monitoring the course of Brucella infection with qPCR-based detection

Che

Bao

et al. 2019

International Journal of Infectious Diseases

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To determine blood Brucella DNA loads between brucellosis patients and those without brucellosis. Methods: The patient group included 350 brucellosis patients. The control was composed of 200 subjects without brucellosis. The extracted DNA from blood was tested by quantitative polymerase chain reaction (qPCR). The cutoff value was determined by receiver operating characteristic curve analysis. A portion of the brucellosis patients were monitored by qPCR during therapy. Results: The detection limit of qPCR was between 1E + 01 cfu/mL and 1E + 08 cfu/mL. The standard curve R 2 reached 0.998. The cutoff value was 4E + 01 cfu/mL, which was determined by comparison of the patient group and the control. The qPCR assay had a specificity of 100% and a sensitivity of 93.14%. The monitoring results showed that the Brucella DNA load decreased in most patients during the first 4 weeks of treatment. One patient with bad treatment compliance showed a rebound. Conclusions: The qPCR results were in accordance with the course of brucellosis in the clinic. The DNA load often reflects the situation of the Brucella-infected patient. The cutoff value provides an important reference of infection. This qPCR-based method can be used to assist in the diagnosis of brucellosis and to adjust the therapy.

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“…As with culture, PCR is considered to have high specificity and has proven to be effective in detecting B. canis in the blood of infected dogs. 3 This assertion, in addition to the agreement of IHC results in 3 of 4 cases, led to the consideration of the dogs in our study that tested positively for B. canis via rtPCR as infected. Although the IHC protocol that we used did not show cross-reaction in cases of E. coli epididymitis in dogs, the polyclonal nature of the antibody allows the possibility for cross-reaction with other gram-negative organisms.…”

mentioning

confidence: 63%

Characterization of epididymal and testicular histologic lesions and use of immunohistochemistry and PCR on formalin-fixed tissues to detectBrucella canisin male dogs

et al. 2021

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In male dogs, Brucella canis frequently causes epididymitis, ultimately resulting in testicular atrophy and infertility. Although B. canis predominantly affects the epididymis, the misleading term “orchitis” is still commonly used by clinicians. Of additional concern, diagnosis in dogs remains challenging because of variable sensitivity and specificity of serologic assays and fluctuations in bacteremia levels in infected dogs, reducing the sensitivity of blood culture. We describe here the histologic lesions in the scrotal contents of 8 dogs suspected of being infected with B. canis and clinically diagnosed with orchitis. We explored the possibility of using immunohistochemistry (IHC) and real-time PCR (rtPCR) in formalin-fixed, paraffin-embedded (FFPE) tissues to detect the presence of B. canis. Epididymitis of variable chronicity was identified in all 8 dogs, with only 3 also exhibiting orchitis. Using rtPCR, the presence of B. canis was identified in 4 of 8 dogs, with 3 of these 4 dogs also positive by IHC. These results suggest that rtPCR and IHC are promising techniques that can be used in FFPE tissues to detect B. canis when other detection techniques are unavailable. Additionally, accurate recognition of epididymitis rather than orchitis in suspect cases could aid in accurate diagnosis.

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Comparison of four polymerase chain reaction assays for the detection of Brucella spp. in clinical samples from dogs

Cited by 15 publications

References 26 publications

Canine Brucellosis: Insights Into the Epidemiologic Situation in Europe

Canine Brucellosis: Insights Into the Epidemiologic Situation in Europe

Monitoring the course of Brucella infection with qPCR-based detection

Characterization of epididymal and testicular histologic lesions and use of immunohistochemistry and PCR on formalin-fixed tissues to detectBrucella canisin male dogs

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