JVMBBS - 2021 - Vol. 7 - iss. 1–2

Journal for Veterinary Medicine, Biotechnology and Biosafety

Volume 7, Issue 1–2, June 2021, Pages 17–20

ISSN 2411-3174 (print version) ISSN 2411-0388 (online version)

DEVELOPMENT OF A REAL-TIME PCR ASSAY FOR THE DETECTION OF BRUCELLA OVIS DNA IN CLINICAL SAMPLES

Marchenko N. V., Lymanska O. Yu., Gerilovych A. P., Bolotin V. I.

National Scientific Center ‘Institute of Experimental and Clinical Veterinary Medicine’, Kharkiv, Ukraine, e-mail: marcenkonata@gmail.com

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Citation for print version: Marchenko, N. V., Lymanska, O. Yu., Gerilovych, A. P. and Bolotin, V. I. (2021) ‘Development of a real-time PCR assay for the detection of Brucella ovis DNA in clinical samples’, Journal for Veterinary Medicine, Biotechnology and Biosafety, 7(1–2), pp. 17–20.

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Citation for online version: Marchenko, N. V., Lymanska, O. Yu., Gerilovych, A. P. and Bolotin, V. I. (2021) ‘Development of a real-time PCR assay for the detection of Brucella ovis DNA in clinical samples’, Journal for Veterinary Medicine, Biotechnology and Biosafety. [Online] 7(1–2), pp. 17–20. DOI: 10.36016/JVMBBS-2021-7-1-2-3.

Summary. The etiological agent of infectious ovine epididymitis is Brucella ovis and for its direct indication in clinical samples several PCR protocols are proposed. This study describes a design and selection of the oligonucleotides for real-time PCR targeting conservative BOV_A0504 gene. The specificity of a real-time PCR was validated using 25 B. ovis field isolates and 14 microorganisms of closely related species. The detection limit of B. ovis in bacterial culture was determined as 3.5×10¹ CFU/mL with Ct value of 37.8. There are no detectable fluorescence signals in the clinical samples from intact animals, whereas bacteriologically confirmed material such as urine and testicle tissue samples were positive. It confirms that the assay is highly specific for detection of B. ovis DNA. Thus, the proposed real-time PCR assay enables fast detection and quantification of B. ovis in clinical material, which can be used as additional test for estimation of the health status of a sheep herd

Keywords: specific primers, infectious ovine epididymitis, sheep

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