Optimization of polymerase chain reaction for the identification of Roe deer, Saiga, and Siberian stag living in Kazakhstan

Vet World Vol.15 August-2022 Article-21

Research Article

Veterinary World, 15(8): 2067-2071

https://doi.org/10.14202/vetworld.2022.2067-2071

Kanatbek Mukantayev¹, Darkhan Kanayev¹, Sholpan Zhumabekova¹, Alexander Shevtsov², Kanat Tursunov¹, Kasim Mukanov², and Yerlan Ramankulov²

1. Laboratory of Immunochemistry and Immunobiotechnology, National Center for Biotechnology, Nur-Sultan, Kazakhstan.
2. Laboratory of Applied Genetics, National Center for Biotechnology, Nur-Sultan, Kazakhstan.

Background and Aim: One of the reasons for the decline in the number of wild species of artiodactyls is poaching and the illegal trading of animal products. Molecular genetic identification of animals from a biological sample effectively proves poaching cases and illegal trade of animal products. This study aimed to develop a polymerase chain reaction (PCR) test that allows for species identification of artiodactyl animals that are most often subject to poaching.

Materials and Methods: Genomic DNA was extracted from meat and blood samples of animals killed by poachers using commercial kits. Three pairs of primers were designed and used to amplify the cytochrome b gene fragment of Roe deer, Saiga antelope, and Siberian stag.

Results: The proposed protocol allows amplification of specific PCR products of 542 bp with Roe deer DNA, 587 bp with Saiga DNA, and 525 bp with Siberian stag DNA. Specificity analysis showed no cross activity with DNA from other animal species. The detection limit of PCR ranged from 15.6 pg to 1.9 pg of DNA in 25 μL of the reaction mixture.

Conclusion: Sequencing the amplified products and subsequent comparison with the corresponding reference sequence showed a similarity ranging from 99.99% to 100%. The PCR based on the developed primers demonstrated high sensitivity and specificity when using DNA from homogeneous and heterogeneous animals. Keywords: Argali, cytochrome b, identification, Roe deer, Saiga, Siberian stag.

Keywords: Argali, cytochrome b, identification, Roe deer, Saiga, Siberian stag.

How to cite this article: Mukantayev K, Kanayev D, Zhumabekova S, Shevtsov A, Tursunov K, Mukanov K, and Ramankulov Y (2022) Optimization of polymerase chain reaction for the identification of Roe deer, Saiga, and Siberian stag living in Kazakhstan, Veterinary World, 15(8): 2067–2071.

Received: 21-04-2022 Accepted: 20-07-2022 Published online: 26-08-2022

Corresponding author: Kanat Tursunov E-mail: kanat_tka@mail.ru

DOI: 10.14202/vetworld.2022.2067-2071

Copyright: Mukantayev, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.