Canine-Specific PCR Assay Targeting Cytochrome b Gene for the Detection of Dog Meat Adulteration in Commercial Frankfurters

Abstract

This report described a cytochrome b (cytb)-based polymerase chain reaction (PCR) assay for the detection of canine tissues in commercial frankfurters. Discriminating detection of canine derivatives in processed food products has important application in halal authentication as well as in health, religions, and fare trades. The assay based on a pair of canine-specific primers that targeted a 100 bp region of canine mithochondrial-cytb gene which is present in multiple copies and highly conserved within the same species. The specificity of the assay was tested against dog and eight most common animal meat species as well as five plant species commonly found in frankfurter formulation. The stability and specificity of the assay were verified under different thermal processing conditions under pure and complex matrices. Three commercial brands of chicken and beef frankfurters were tested in triplicate, and specific PCR products were obtained only from deliberately contaminated formulations. The detection limit of the assay was 0.1 % (0.02 ng DNA) of canine meat spiked with other meats in a typical frankfurter formulation. Shorter amplicon length, superior stability, and higher sensitivity of the assay suggested its potential application in the screening of canine-origin biomaterials in processed food products.

Lab-on-a-Chip PCR-RFLP Assay for the Detection of Canine DNA in Burger Formulations

“…Canine Species Specificity Species-specific PCR assay is a commonly used technique for species authentication in raw and processed foods (Ali et al 2013;Mane et al 2009;Rahman et al 2014). The presence of a single mismatch in the primer binding site may reduce the efficiency of PCR assay or may lead to PCR amplification failure (Wu et al 2009).…”

“…In c, Analysis of the primer mismatch and pairwise distance using 3D plot commercial food processing conditions, causing failure or truncated PCR products. We used here shorter DNA target (100 bp) and found that it was stable under various food processing treatments (Ali et al 2013;Rahman et al 2014). …”

“…The assay was less sensitive than the present one in terms of the amount of template DNA used (125 vs. 20 ng) and the limit of detection (0.1 vs 0.01 %). Recently for dog meat detection, Ali et al (2013) documented a cytb-based PCR assay using agarose gel end-point detection technique with a sensitivity of 0.1 % (w/w) (0.02 ng DNA). Compared to this, CE-based end-point detection scheme of the present assay provided a higher sensitivity of 0.01 % (w/w) (0.002 ng canine DNA) using similar admixtures of dog meat with chicken and beef.…”

“…Furthermore, availability of stray dogs in certain countries (Kumarapeli and Awerbuch-Friedlander 2009;Totton et al 2010) has made dog meat a potential adulterant in costly meats such as beef, lamb, and chicken. Henceforth, several PCR assays for canine species detection have been documented (Abdel-Rahman et al 2009;Abdulmawjood et al 2003;Ali et al 2013;Arslan et al 2006;Gao et al 2004;İlhak and Arslan 2007;Martín et al 2007;Rahman et al 2014). However, all of these assays are based on traditional agarose-gel electrophoresis for PCR endpoint detection, and none of them have been tested in burger formulation.…”

Application of FTIR Spectroscopy and Chemometrics for Halal Authentication of Beef Meatball Adulterated with Dog Meat

“…Rahman et al [5] have used PCR assay using cytochrome b gene as a target for detecting DM adulteration in meatball formulation. PCR assay using specific-specific primer on cytochrome b was also used for authentication of commercial Frankfurters from DM [12]. PCR using specific primer and restriction fragment length polymorphism (RFLP) was used for identification of meat sourced from cat, dog, donkey and horse [13].…”