Quantitative real‐time PCR assays to detect DNA degradation in soy‐based food products

Abstract

AbstractBACKGROUND: Polymerase chain reaction (PCR)‐based DNA diagnostics have grown in importance for assessing food quality and safety. PCR diagnostic reliability and sensitivity depend on the quality of the DNA extractions, including the extent of DNA degradation and the presence of PCR inhibitors.RESULTS: An approach has been described that quantifies the extent of DNA degradation in soy food samples using anchored real‐time PCR (qPCR) assays that amplify target sequences ranging from < 100 to > 1000 bp, based on two endogenous soy sequences. DNA degradation was quantified for model foods produced in the laboratory (cooked soy meal, tofu) as well as purchased soy‐containing food products. Considerably less than 1% of the total DNA extracted from samples was available for amplification of the longest amplicons (830 and 1022 bp) from the most highly processed food products (e.g., soy‐based infant formula).CONCLUSION: The utility of anchored qPCR assays was demonstrated for characterizing the amount of DNA that is available for amplifying different‐length PCR products from a range of soy‐containing processed food products. This approach should be useful for estimating the amount of amplifiable DNA in food ingredients in cases where food processing has caused degradation of DNA. Copyright © 2009 Society of Chemical Industry