Issue 1

Journal for Veterinary Medicine, Biotechnology and Biosafety

Volume 1, Issue 1, March 2015, Pages 15–22

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


Stegniy B. T., Goraichuk I. V., Gerilovych A. P., Kucheryavenko R. O., Bolotin V. I., Solodiankin O. S.

National Scientific Center “Institute of xperimental and Clinical Veterinary Medicine”, Kharkiv, Ukraine, e-mail:,

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Citation for print version: Stegniy, B. T., Goraichuk, I. V., Gerilovych, A. P., Kucheryavenko, R. O., Bolotin, V. I. and Solodiankin, O. S. (2015) ‘Creation molecular-genetic control system of Pestivirus contamination in biotechnology objects’, Journal for Veterinary Medicine, Biotechnology and Biosafety, 1(1), pp. 15–22.

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Citation for online version: Stegniy, B. T., Goraichuk, I. V., Gerilovych, A. P., Kucheryavenko, R. O., Bolotin, V. I. and Solodiankin, O. S. (2015) ‘Creation molecular-genetic control system of Pestivirus contamination in biotechnology objects’, Journal for Veterinary Medicine, Biotechnology and Biosafety. [Online] 1(1), pp. 15–22. Available at:

Summary. This study aimed on (i) creation molecular-genetic control system of pestivirus contamination in biotechnology objects, (ii) identification of persistently infected with bovine viral diarrhea virus (BVDV) animals and (iii) genetic typing of selected BVDV isolates.

RNA extraction, cloning, polymerase chain reaction (PCR), real-time PCR, enzyme-linked immunosorbent assay, serum neutralization test, sequencing.

It was shown that we had constructed the recombinant plasmids with insertion Erns gene fragment (826 base pair) of BVDV-1 and BVDV-2. Also we had developed and optimized parameters of duplex PCR for the simultaneous indication Mollicutes DNA and BVDV RNA, with the possibility of nested PCR for further identification of BVDV genotypes. Specific BVDV antibodies were detected in 725 of 1042 (69.6 %) analyzed samples. In this study 5 persistently infected with BVDV animals were detected in farms B and C of Kharkiv region. The genetic typing of viral isolates revealed that only BVDV-1 viruses were present. The phylogenetic analysis confirmed two BVDV-1 subtypes, namely b and f and revealed that all viruses from the farm B of Kharkiv region and from biotechnological objects were typed as BVDV-1b, but virus from the farm C of Kharkiv region and farm of Kherson region were typed as BVDV-1f.

The obtained recombinant plasmids can be used as a positive control for PCR and test-system for control of pestivirus contamination in biotechnology objects. Our results indicated that the BVDV infection is widespread in cattle herds in the eastern Ukraine, that requires further applying of new approaches to improve the current situation.

Keywords: bovine viral diarrhea virus, pestivirus contamination, cloning, pTZ57R/T, restriction enzyme digestion analysis, ELISA, SNT, PCR, real-time PCR, genotyping, phylogenetic analysis.


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