Evaluation of real-time PCR assays for detection and quantification of fraudulent addition of bovine milk to caprine and ovine milk for cheese manufacture

Many typical Italian cheeses made from ovine milk are certified as Protected Designation of Origin (PDO). Because caprine and ovine milk production is limited, the fraudulent addition of cows' milk is widespread. In addition, some compounds in bovine milk have high allergenic potential; therefore, such fraud also has implications for consumer health. In this study, a real-time polymerase chain reaction (real-time PCR) test was developed to detect and quantify cow's milk in caprine and ovine cheeses, based on two target genes. The mitochondrial Cytochrome b gene (Cytb) of Bos taurus was used to detect and quantify bovine DNA. The nuclear gene myostatin (Myo), nuclear ribosomal gene 18S, or mitochondrial gene 16S were used alternatively as universal reference markers. Caprine (n = 30) and ovine (n = 51) cheese samples were purchased and analyzed and most were shown to be contaminated by bovine milk. Pairwise analysis of quantification data using a Spearmann Rank Correlation test demonstrated a highly significant correlation between data obtained with the different reference assays.