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

Abstract

Canine species detection in foods is important in the perspectives of health, religions, and fare-trade food business. This study describes a very short-amplicon length Polymerase Chain Reaction (PCR)-Restriction Fragment Length Polymorphism (RFLP) assay with lab-on-a-chip detection platform for the authentication of canine DNA in processed foods. A 100-bp fragment of canine mitochondrial Cytochrome b (cytb) gene was selected and amplified using a pair of canine-specific primers. The amplified PCR products were validated by RFLP analysis using lab-on-a-chip microfluidic bioanalyzer kit. Both gel-image and electropherograms authenticated the canine-specific PCR products before (100 bp) and after restriction digestion (51, 30, and 19 bp). The assay successfully detected 0.0001-ng canine DNA under pure state and 0.01 % (w/w) canine meat spiked in chicken and beef burger formulations. Screening of eight commercial burgers across Malaysia did not reveal any canine adulteration. We believe the assay would find potential applications in food industries, Halal food regulatory bodies and animal right protection authorities across the globe.

Double gene targeting multiplex PCR-RFLP detects Crocodylus porosus in chicken meatball and traditional medicine

“…[19] has carried out PCR-RFLP to discriminate C. porosus, C. palustris and C. gangeticus in fresh blood samples, nonetheless, the assay was not tested under food processing conditions. Previously, successful PCR-RFLP assay has been carried out in various food products, namely frankfurter, [27,42] meatballs, [34] burgers [29,41,43] for the detection of rabbit, rat, squirrel, beef, buffalo, pig, cat, dog, and monkey.…”

Lab-on-a-chip-based PCR-RFLP assay for the confirmed detection of short-length feline DNA in food

“…Feline PCR product was amplified from all contaminated meatballs, showing the tested LOD of the assay was 0.01% (w/w) feline meat. Previously, Ali et al (2013) and Rahman et al (2015) detected 0.1% (w/w) and 0.01% (w/w) canine DNA in frankfurters and burgers formulations, respectively. However, we do not think there is a need for adulteration detection below 0.01% since adulteration at this level is not profitable and unlikely to be done.…”

Advances in Authenticity Testing for Meat Speciation

“…Table 14.2 summarizes the most relevant information from reported PCR-RFLP methods, from which it can be inferred that mtDNA markers are mostly targeted to differentiate domestic and game meat species. Most PCR-RFLP methods use gel More recently, chip-based capillary electrophoresis has emerged as an alternative to conventional electrophoresis for RFLP analysis for meat authentication, producing higher resolution band patterns, shorter analysis times, higher versatility, reproducibility, and suitability for automation (Ali et al, 2012a(Ali et al, , 2015aFajardo et al, 2009a;Rahman et al, 2015). …”