Estri Laras Arumingtyas, Joni Kusnadi, Muhammad Thoriq Farizi, Ainun Sayyidah Zakiyah, Galuh Arkana
Background and Objective: Buffalo plays a role in providing animal protein in Indonesia. Their meat and skins are widely used as raw materials for processed foods. However, the high demand for buffalo products is not proportionate with the current supply; thus, creating chances for food fabricating. This practice is not only illegal but also causes serious risks to food safety and halal compliance. The detection of adulteration in food product can be carried out using polymerase chain reaction technique and specific primers that target buffalo DNA. This research aimed to develop primers that could accurately identify buffalo DNA. Material and Methods: The primers were designed based on the sequence of the buffalo cytochrome b gene. The effectiveness of these primers in recognizing and amplifying buffalo DNA was assessed using polymerase chain reaction technique. Specificity assessments were carried out to assess the primer capability to detect buffalo DNA, with comparative controls including cattle, goat, chicken, pig, dog and mouse DNA. Sensitivity assessments were carried out to assess the minimum DNA concentration detectable by polymerase chain reaction. Then, the ability of these primers in identifying buffalo skin crackers was assessed against controls of cow skin crackers, goat skin crackers, and crispy chicken skin and pork skin crackers.Results and Conclusion: The polymerase chain reaction results indicated that the Buffalo_5.1 primer pair (forward 5’-TTAGTACTATTCGCACCCGACCTC-3’ and reverse 5’-TCGTTGTTTGGATGTATGTAGCAG-3’) successfully amplified buffalo DNA specifically, with a detection limit of up to 10-3 ng μl-1 (assuming that the solution included a density of 1 g ml-1 equal to 10-⁷ % w/w), which could potentially increase to 10-5 ng μl-1 (10-⁷ % w/w) under optimal conditions. Furthermore, it was able to detect buffalo DNA in buffalo-skin cracker products even at low DNA purity levels. These results suggest that the Buffalo_5.1 primer includes the potential to serve as a reliable molecular marker in polymerase chain reaction-based food authentication studies. This open-access article distributed under the terms of the Creative Commons Attribution Non Commercial 4.0 License (CC BY-NC 4.0).To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Brawijaya, Indonesia; Department of Food Science and Technology, Faculty of Agricultural Technology, Universitas Brawijaya, Indonesia
