Development of rapid PCR-RFLP technique for identification of sheep, cattle and goat’s species and fraud detection in Iranian commercial meat products

Abstract

Identification of animal species used in commercial meat products is important with respect to economic and sanitary issues. The aim of this research was to realize ruminant species in meat products using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. The universal CB7u primers pair was used for amplifying ~195 bp fragment from a variable region of cytochrome-b mitochondrial DNA gene by polymerase chain reaction. Species differentiation was realized by digestion of the amplified ~195 bp fragments with Sse9I restriction enzyme. The results indicate that 7/7 of Kebab loghmeh, 9/10 of minced meat, 4/8 of beef burger and 2/5 samples of canned stew samples, were contaminated with one of prohibited ruminant species residual. Furthermore, the results reveal that 5/30 of samples had cross-contamination with a mixture of meat originated from various species, which was against the labelled nutrition information. Our results indicate that the PCR-RFLP technique is a powerful and reproducible test for detection and separation of ruminant species residuals in commercial meat products, especially in developing countries.

Keywords: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), mitochondrial DNA, ruminant species, commercial meat products, cytochrome-b gene