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Journal for Veterinary Medicine, Biotechnology and Biosafety

Volume 7, Issue 3, September 2021, Pages 19–23

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

DEVELOPMENT OF RECOMBINANT POSITIVE CONTROL FOR DETECTION OF PORCINE CIRCOVIRUS TYPE 3 BY POLYMERASE CHAIN REACTION

Rudova N. G., Lymanska О. Yu., Bolotin V. I., Stegniy B. T., Solodiankin О. S., Gerilovych А. P.

National Scientific Center ‘Institute of Experimental and Clinical Veterinary Medicine’, Kharkiv, Ukraine, e-mail: rudovanatawa@ukr.net

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Citation for print version: Rudova, N. G., Lymanska, О. Yu., Bolotin, V. I., Stegniy, B. T., Solodiankin, О. S. and Gerilovych, А. P. (2021) ‘Development of recombinant positive control for detection of porcine circovirus type 3 by polymerase chain reaction’, Journal for Veterinary Medicine, Biotechnology and Biosafety, 7(3), pp. 19–23.

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Citation for online version: Rudova, N. G., Lymanska, О. Yu., Bolotin, V. I., Stegniy, B. T., Solodiankin, О. S. and Gerilovych, А. P. (2021) ‘Development of recombinant positive control for detection of porcine circovirus type 3 by polymerase chain reaction’, Journal for Veterinary Medicine, Biotechnology and Biosafety. [Online] 7(3), pp. 19–23. DOI: 10.36016/JVMBBS-2021-7-3-3.

Summary. This work aimed to obtain positive control using recombinant DNA technology for detection by PCR of a new poorly studied pathogen — porcine circovirus type 3. Recombinant positive control was designed using Clone Manager Basic. As a vector in the creation of recombinant control we used plasmid pTZ57R/T, as an insert — a fragment of the gene rep PCV-3 with the length of 418 nucleotide pairs, obtained by classical PCR. Transformation of competent cells of E. coli strain DH5a was carried out by chemical poration, followed by plating on LB-medium with the addition of ampicillin at a final concentration of 100 μg/ml. The selection of E. coli cell colonies was performed by the marker of antibiotic resistance to ampicillin. The presence of a specific insert was checked by PCR with electrophoretic visualization of the results. The developed recombinant positive control can be used for the monitoring of biological samples from pigs for the presence of genetic material PCV-3 using molecular technologies

Keywords: PCV-3, PCR, plasmid pTZ57R/T, gene rep, E. coli strain DH5a

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