A Fast and Reliable Real-Time PCR Method for Detection of Ten Animal Species in Meat Products.

Species substitution in meat products is a common problem reported worldwide. This type of food fraud is, typically, an intentional act for economic gain, using sources of low-priced meats in high-value meat products. Consequences include economic, health, and religious concerns. Highly sensitive and efficient techniques are thus required to detect meat species. This paper describes a method based on real-time PCR to detect 10 animal species (Bos taurus, Sus scrofa, Ovis aries, Capra hircus, Gallus gallus, Meleagris gallopavo, Bubalus bubalis, Equus caballus, Felis catus, and Canis familiaris) in meat product. The method combines species-specific and universal (used here as internal positive control) primers, and applies melt curve analysis for amplicon checking. Method accuracy was evaluated on 46 experimental meat mixtures and all species were correctly identified in all cases, at 1% test sensitivity. Analysis of 14 commercial meat products revealed that 6 of 14 samples had nondeclared bovine and/or chicken material. We performed an interlaboratory comparison using the reference meat mixtures and commercial samples, achieving 100% of reproducibility. The developed test proved to be effective and reliable for routine analysis of meat products. PRACTICAL APPLICATION: This paper describes a fast and reliable method for species detection in meat products based on real-time PCR. It can be applied for analysis of in natura or processed meat. The method proposed here can play an important role in controlling the origin of meat products, ensuring their quality and safety for the entire food industry-producers to consumers.