Detection of Adulteration in Italian Mozzarella Cheese Using Mitochondrial DNA Templates as Biomarkers

Considering the importance of monitoring adulterations of genuine cheeses in the dairy industry, a polymerase chain reaction–based method was developed to detect bovine-specific mitochondrial DNA sequence in Italian water buffalo Mozzarella cheese. DNA was isolated from cheese matrix and governing liquid by organic extractions and kit purifications. Amplifications of a 134-bp fragment were performed with a bovine–specific set of primers designed on the sequence alignment of bovine and buffalo mitochondrial cytochrome oxidase subunit I. The specificity of the primers was tested using DNA from the blood of two species (water buffalo and bovine), which are present together in adulterated Italian Mozzarella cheese. This method reliably detected a content of 0.5 % of bovine milk, making it suitable for routine fraud monitoring.