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

TLDR

Real‑time PCR assays for detecting meat in processed products have focused on small amplicons, yet the relationship between amplification rates and amplicon size in such products remains unclear. The study aimed to investigate how amplification rates vary with amplicon size in processed meat products. Real‑time PCR assays with a range of amplicon sizes were applied to processed meat matrices to assess amplification rates. The most sensitive assays used the smallest amplicons, but amplification was still detectable with 351‑bp amplicons in highly processed samples, showing that larger amplicons can amplify efficiently and should be chosen only when specificity is not compromised.

Abstract

The design of real-time polymerase chain reaction (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 rates and the 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 rates, in relation to amplicon size, in processed meat matrices. Although the most sensitive assays were those that used the smallest amplicons, amplification was still observed using amplicons of 351 base pairs for highly processed samples. 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.