Development of real-time polymerase chain reaction for analysis of rat meat ( Bandicota bengalensis ) in beef meatballs for halal authentication.

Autor: Rohman A; Faculty of Pharmacy, Universitas Gadjah Mada, Yogyakarta, Indonesia.; Halal Center, Gadjah Mada University, Yogyakarta, Indonesia., Nawwaruddin HH; Faculty of Pharmacy, Universitas Gadjah Mada, Yogyakarta, Indonesia.; Halal Center, Gadjah Mada University, Yogyakarta, Indonesia., Hossain MAM; Nanotechnology and Catalysis Research Centre (NANOCAT), University of Malaya, Kuala Lumpur, Malaysia., Laksitorini MD; Faculty of Pharmacy, Universitas Gadjah Mada, Yogyakarta, Indonesia.; Halal Center, Gadjah Mada University, Yogyakarta, Indonesia., Lestari D; Faculty of Pharmacy, Universitas Muhammadiyah Kalimantan Timur, Samarinda, Indonesia.
Jazyk: angličtina
Zdroj: Open veterinary journal [Open Vet J] 2024 Sep; Vol. 14 (9), pp. 2484-2492. Date of Electronic Publication: 2024 Sep 30.
DOI: 10.5455/OVJ.2024.v14.i9.37
Abstrakt: Background: Consumer awareness of food adulteration is increasing nowadays. Motivated by economic gain, unethical meat producers try to blend halal meat such as beef with non-halal meat like rat meat (RM).
Aim: This study aims to develop a real-time polymerase chain reaction (RT-PCR) analysis method to analyze the presence of RM in beef meatballs.
Methods: This research was carried out in the following stages: primer design, DNA isolation, analysis of DNA isolates, the optimization of primer annealing temperature, primer specificity test, sensitivity, and repeatability. The validated RT-PCR method was then used to analyze the marketed meatball samples.
Results: The result showed that the designed primer targeting on ND2 gene set rat mt-DNA (forward: ACTCCATATCTCTCACCATATTTCC; reverse: GGGTTAGGGTACTTAGGATTGTTAG), had good specificity at an optimal annealing temperature of 56.3 o C over the other eight species. The developed RT-PCR method produces a limit detection value of 195.31 pg, coefficient of determination ( R 2 ) for linearity of 0.983, amplification efficiency (E) of 100%, and CV value for amplification response of 1.8%. The result showed that the developed RT-PCR method did not detect the presence of RM DNA in eight marketed beef meatball samples.
Conclusion: The developed method meets the acceptance criteria for RT-PCR and can be used as a halal authentication method to identify the presence of RM in beef meatballs.
Competing Interests: The authors state no conflict of interest.
Databáze: MEDLINE
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