Short Amplicon-Length PCR Assay Targeting Mitochondrial Cytochrome b Gene for the Detection of Feline Meats in Burger Formulation
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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.
- Abdel-Rahman S, El-Saadani M, Ashry K, Haggag A (2009) Detection of adulteration and identification of cat’s, dog’s, donkey’s and horse’s meat using species-specific PCR and PCR-RFLP techniques. Aust J Basic Appl Sci 3(3):1716–1719Google Scholar
- Adams DS (2013) Lab math: a handbook of measurements, calculations, and other quantitative skills for use at the bench, 2nd edn. Cold Spring Harbor 361 Laboratory Press, New York 11724, United States, p 323Google Scholar
- Ali ME, Hashim U, Mustafa S, Che Man YB (2012b) Swine-specific PCR-RFLP assay targeting mitochondrial cytochrome B gene for semiquantitative detection of pork in commercial meat products. Food Anal Methods 5:613–623Google Scholar
- Ali ME, Mustafa S, Hashim U, Man Y, Foo K (2012d) Nanobioprobe for the determination of pork adulteration in burger formulations. J Nanomater 2012, 8Google Scholar
- Ali ME, Rahman MM, Hamid SBA, Mustafa S, Bhassu S, Hashim U (2013) Canine-specific pcr assay targeting cytochrome b gene for the detection of dog meat adulteration in commercial frankfurters. Food Anal Methods 1–8Google Scholar
- Ali ME, Razzak MA, Hamid SBA (2014) Multiplex PCR in species authentication: probability and prospects—a review. Food Anal Methods 1–17Google Scholar
- Anitei S (2006) The origin of SARS epidemics found in civet cat meat consumption in Southern China.Retrieved 4 November 2014 http://news.softpedia.com/news/The-Origin-of-SARS-Epidemics-Found-in-Civet-Cat-Meat-Consumption-in-Southern-China-40897.shtml
- Anonymous (2014) Malaysia: marketing to Muslim consumers. Retrieved 20 November 2014 http://economists-pick-research.hktdc.com/business-news/article/Research Articles/Malaysia-marketing-to-Muslim consumers/rp/en/1/1X000000/1X09XTD8.htm
- Chatterji A (2013) Dog or cat meat suspected in mystery Indian ‘lamb’ curry. International Buisness Times. Retrieved fromhttp://www.ibtimes.co.uk/dog-cat-meat-horse-india-lamb-curry-451343
- Commission E. (2002) Commission Decision 2002/657/EC of 12 August 2002 implementing Council Directive 96/23/EC concerning the performance of analytical methods and the interpretation of results. Off J Eur Comm L 221:8–36Google Scholar
- Doosti A, Ghasemi Dehkordi P, Rahimi E (2011) Molecular assay to fraud identification of meat products. J Food Sci Technol 1-5Google Scholar
- İlhak Oİ, Arslan A (2007) Identification of meat species by polymerase chain reaction (PCR) techniques. Turk J Vet Anim Sci 31:159–163Google Scholar
- Khattak JZK, Mir A, Anwar Z, Wahedi HM, Abbas G, Khattak HZK, Ismatullah H (2011) Concept of halal food and biotechnology. Adv J Food Sci Technol 3(5):385–389Google Scholar
- Ong S, Cheah YK, Robin T, Wolmon Gunsalam J, Mat Isaa Z, Chai LC, Yuli H, Mohamad Ghazali F, Radu S (2007) Meat molecular detection: sensitivity of polymerase chain reaction-restriction fragment length polymorphism in species differentiation of meat from animal origin. ASEAN Food J 14:51–59Google Scholar
- Phillips T (2013) Chinese police find slaughterhouse selling cat meat. The Telegraph. Retreived from http://www.telegraph.co.uk/news/worldnews/asia/china/10417032/Chinese-police-find-slaughterhouse-selling-cat-meat.html
- Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406Google Scholar
- Salama (2013) “Global halal traders gather in malaysia amid muslim population growth.” Retrieved 10 January, 2014, from http://halalfocus.net/global-halal-traders-gather-in-malaysia-amid-muslim-population-growth/
- Tobe SS, Linacre AM (2008) A multiplex assay to identify 18 European mammal species from mixtures using the mitochondrial cytochrome b gene. Electrophoresis 29:340–347. doi: 10.1002/elps.200700706