Short Amplicon-Length PCR Assay Targeting Mitochondrial Cytochrome b Gene for the Detection of Feline Meats in Burger Formulation
Consumption or mixing of feline ingredients in halal and kosher foods is forbidden, and various diseases such as SARS, anthrax, and hepatitis could be transmitted through feline meats. However, since feline species are abundant across the world without any market price and their meats are consumed in exotic foods, the chances of their adulteration in common meats are very high. Several recent reports appreciated short amplicon-length PCR assays for species authentication in processed foods assuming that shorter targets would be thermodynamically more stable than longer ones under natural decomposition and food processing treatments. However, scientific evidence to prove this hypothesis is rarely documented. For the first time, we developed here a PCR assay targeting only a 69-bp site of feline mitochondrial cytochrome b gene, and its authenticity was confirmed by AluI restriction enzyme followed by its separation and detection on a lab-on-a-chip-based automated electrophoretic system. The exceptional target stability was systematically proven over the previously documented shortest target (108 bp) under extreme autoclaving and microwaving treatments both in pure and mixed matrices. The assay specificity was tested against 14 terrestrial and aquatic species commonly consumed in foods, and no cross-species detection was observed. The limit of detection of the assay was 0.1 pg of feline DNA and 0.01 % (w/w) of feline meats in raw meats and cooked burgers, respectively.
KeywordsFeline ingredients Short amplicon-length PCR assay Burger formulation Compromised states Lab-on-a-chip
M.A. Amin is a recipient of Graduate Research Assistantship (GRA) from the University of Malaya, Kuala Lumpur, Malaysia. M.A. Amin was paid by “AkaunPengrusanCombicat no. 31377” and “AkaunPenyelidikan Flagship-RU002-2014” to S.B.A. Hamid, and consumables were paid by GC001A-14SBS to M.E. Ali.
Compliance with Ethics Requirements
Ethical clearance of ref. no: NANOCAT 26/09/3013/ MAA (R) was obtained from the Institutional Animal Care and Use Committee, University of Malaya (UM IACUC), and all experiments were conducted following the national and institutional guideline while handling animal meats used in this study.
Conflict of Interest
Md. Eaqub Ali declares that has supervised this work, provided intellectual supports, and extensively edited the manuscript, and he has no conflict of interest to publish this paper. Md. Al Amin declares that he performed this research and drafted manuscript, and he has no conflict of interest to publish this paper. Md. Abdur Razzak declares that he helped Md. Al Amin in sample collection, data analysis, and preparing manuscript, and he has no conflict of interest to publish this paper. Sharifah Bee Abd Hamid declares that she provided financial and logistic supports to carry out this research, and she has no conflict of interest to publish this paper. Md. Mahfujur Rahman declares that he helped Md. Al Amin in data analysis, and he has no conflict of interest to publish this paper. NurRaifana Abdul Rashid declares that she helped Md. Al Amin in manuscript drafting, and she has no conflict of interest to publish this paper. Asing declares that he helped Md. Al Amin in experimental section, and he has no conflict of interest to publish this paper.
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