SPECIES IDENTIFICATION USING POLYMERASE CHAIN REACTION-RESTRICTION FRAGMENT LENGTH POLYMORPHISM (PCR-RFLP) ANALYSIS OF MEAT AND MEAT MIXTURES

Document Type : Research article

Authors

Dept. of Forensic Medicine and Toxicology, Fac. of Vet. Medicine, Assiut University. Assiut

Abstract

In this study we used the Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) technique to differentiate between meat among ten animal species (cattle, buffalo, sheep, pig, dog, horse, donkey, rat, mice and camel). Such discrimination tests may have important applications in the forensic science, agriculture, quarantine and customs fields. Muscle tissue samples were taken from the different animal species (5 each) and kept frozen till used. Meat mixtures were prepared by adding 0.5 or 1.0 % levels of pork, donkey and dog meat to meat samples of cattle (w/w). The 359 bp fragments of cytochrome b (Cytb) gene were generated and distinct digestion patterns of these DNA fragment were observed after HaeIII and Rsal enzymes treatment. The results indicated that PCR amplification yielded 359 bp fragments in each of these species and mixtures. The amplicons were digested with two restriction enzymes resulting in a pattern that could identify and differentiate each of the above species except in camel which need specific restriction enzyme to digest its PCR product. In conclusion, these ten species of animals can be qualitatively identified and differentiated by PCR-RFLP of mitochondrial Cytb with the restriction enzymes. This method can be applied also in meat mixture to identify the adulteration with minute amount of undesirable meat in consumer meat products.

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