Pig Species Identification in Meatballs Using Polymerase Chain Reaction Restriction Fragment Length Polymorphism

Abstract

The information given to consumers is essential for them to choose one food product over another. The falsification of food contents on product labels is a widespread problem, especially with products related with pig or others prohibited food in Islam. Proving conclusively that fraud has occurred requires the detection and quantification of food constituents. Falsifications of meat or food are often biochemically similar to the materials they replace, consequently the identification and measurement extremely difficult. The DNA based methods have now been successfully adapted for detection of food substitution. In this research, Polymerase Chain Reaction (PCR) products of cytochrome b mitochondrial DNA gene were applied to identify the existence of pig in meatball product. Genomic DNA of pig, bovine, and chicken were isolated and subjected to PCR amplification targeting the mitochondrial cytochrome b gene. Pig species differentiation was determined by digestion of obtained 359 bp amplified product with BseDI restriction enzymes, which generated pig species electrophoresis pattern. PCR-Restriction Fragment Length Polymorphism (RFLP) revealed the presence of the pig meat in meatball product and distinguished between bovine, chicken, and pig sample. Pig mitochondrial cytochrome DNA gene was cleaved into 228 bp and 131 bp fragments but the bovine, and chicken cytochrome b gene were not digested by BseDI enzyme. The digestion was conducted at 55oC for 3 h and visualization of the digest product was performed in 2% agarose gel. PCR-RFLP technique using BseDI restriction enzymes is reliable for the detection of the pig meat in meatball for the Halal authentication.