Open Access
Research (Published online: 26-08-2022)
21. Optimization of polymerase chain reaction for the identification of Roe deer, Saiga, and Siberian stag living in Kazakhstan
Kanatbek Mukantayev, Darkhan Kanayev, Sholpan Zhumabekova, Alexander Shevtsov, Kanat Tursunov, Kasim Mukanov, and Yerlan Ramankulov
Veterinary World, 15(8): 2067-2071

Kanatbek Mukantayev: Laboratory of Immunochemistry and Immunobiotechnology, National Center for Biotechnology, Nur-Sultan, Kazakhstan.
Darkhan Kanayev: Laboratory of Immunochemistry and Immunobiotechnology, National Center for Biotechnology, Nur-Sultan, Kazakhstan.
Sholpan Zhumabekova: Laboratory of Immunochemistry and Immunobiotechnology, National Center for Biotechnology, Nur-Sultan, Kazakhstan.
Alexander Shevtsov: Laboratory of Applied Genetics, National Center for Biotechnology, Nur-Sultan, Kazakhstan.
Kanat Tursunov: Laboratory of Immunochemistry and Immunobiotechnology, National Center for Biotechnology, Nur-Sultan, Kazakhstan.
Kasim Mukanov: Laboratory of Applied Genetics, National Center for Biotechnology, Nur-Sultan, Kazakhstan.
Yerlan Ramankulov: Laboratory of Applied Genetics, National Center for Biotechnology, Nur-Sultan, Kazakhstan.

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

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

Corresponding author: Kanat Tursunov

E-mail: kanat_tka@mail.ru

Citation: 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.
Abstract

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.