Effect of heat and pressure processing on DNA fragmentation and implications for the detection of meat using a real-time polymerase chain reaction

Abstract\n The design of real-time PCR assays for the detection of meat in processed products has focused on using small amplicons often to the detriment of specificity. However, the relationship between amplification efficiency rates and amplicon size for processed meat products has yet to be determined. To investigate this relationship real-time PCR assays were designed to give a series of amplicons of increasing size. These assays were then used to assess amplification efficiencyrates, in relation to amplicon size, in processed meat matrices. It was found that, although the most sensitive assays were those which used the smallest amplicons, efficient amplification was still observed using amplicons of 350 351 base pairs for highly processed samples, particularly for chicken and cow. It was found therefore, that although in general, amplicons should be as small as possible, larger amplicons give efficient amplification and that small amplicons should not be chosen if they compromise assay specificity.