Identification of bovine, ovine and caprine pure and binary mixtures of raw and heat processed meats using species specific size markers targeting mitochondrial genome

A specific polymerase chain reaction (PCR) method was applied for identification of bovine (Bos taurus), ovine (Ovis aries) and caprine (Capra hircus) pure and binary mixtures of raw and heat-processed meats. These meats are used in food industry products and/or for direct consumption of consumers. The mitochondrial DNA was amplified as a template in a PCR reaction by use of specific primers related to each species. Specific primers with mitochondrial origin amplified amplicons with the length of 300, 172 and 122 bp in target regions in cattle, sheep and goat, respectively. For determination of the primer sensitivity a set of binary meat mixtures with 0.1, 0.5, 1, 5, 10, 50 and 100% based on weight ratio was tested. The detection limit was found to be 0.1% for all samples. In heat-processing program boiled water for 20 min, 121°C for 20 and 30 min and 127°C for 20 min by autoclaving was used, similar to the conditions that are used in pasteurization, sterilization and also meat processing in industrial factories. The performance of the method was not affected by prolonged heat treatments. The results obtained in the present study confirmed the efficiency of species specific primers for targeting of mitochondrial genome in order to detect bovine, ovine and caprine pure and binary mixtures of raw and heat processed meats with high sensitivity and accuracy.