Short Amplicon-Length PCR Assay Targeting Mitochondrial Cytochrome b Gene for the Detection of Feline Meats in Burger Fo
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Short Amplicon-Length PCR Assay Targeting Mitochondrial Cytochrome b Gene for the Detection of Feline Meats in Burger Formulation Md. Eaqub Ali 1,2 & Md. Al Amin 1 & Md. Abdur Razzak 1 & Sharifah Bee Abd Hamid 1 & Md. Mahfujur Rahman 1 & NurRaifana Abdul Rashid 1 & Asing 1
Received: 27 November 2014 / Accepted: 22 June 2015 # Springer Science+Business Media New York 2015
Abstract 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 %
* Md. Eaqub Ali [email protected] 1
Nanotechnology and Catalysis Research Centre, University of Malaya, Kuala Lumpur 50603, Malaysia
2
Centre for Research in Biotechnology for Agriculture (CEBAR), University of Malaya, Kuala Lumpur, Malaysia
(w/w) of feline meats in raw meats and cooked burgers, respectively. Keywords Feline ingredients . Short amplicon-length PCR assay . Burger formulation . Compromised states . Lab-on-a-chip
Introduction Global halal food markets are rapidly expanding because of its special health and religion compliant attributes (Ali et al. 2014). The current consumers of this market has reached to 1.8 billion (Anonymous 2014) and turnover has crossed US$ 700 billion in 2012 (Salama 2013). Halal or Islamic meats are tastier, tender, healthier, safer, and stay fresh for longer time because of their better resistivity to bacterial invasion given by the specialized slaughtering process that removes blood from carcases (Khattak et al. 2011). Ready-made foods such as burgers, pizzas, hot dogs, sandwiches, soups, cookies, candies, creams, and numerous others have got increasing popularity among the working people and teenagers because of their convenient features and availability in roadside restaurants and groceries (Ali
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