New PCR Real-Time TaqAAan system for beef quantification in meat-based products

摘要

In this study, a PCR Real-Time Tag-Man type System was developed for the rapid and highly specific detection and quantification of the beef present in meat-based products. DNA was isolated using the CTAB modified protocol which is able to quantitatively extract all the species present in the sample thereby supplying a high-quality extract. The primers and the TaqMan probe amplify a fragment of 146 base pairs of the mitochondrial gene coding for Cytochrome b. To finalize the method, a calibration curve was built by mixing calculated amounts of pure beef and pork DNA, and binary beef/pork mixtures were analysed. Tests carried out demonstrated good accuracy in the 0.1-25% beef range. Field tests showed that the method is applicable and equally effective also with complex foods undergoing mild heat treatment.