Factors Influencing the Accuracy of Measurements with Real-Time PCR: The Example of the Determination of Processed Animal Proteins

Quantification of traces of processed animal proteins (PAPs) in feedingstuffs is an important requirement to enforce upcoming European legislation. One of the techniques proposed in this context is real-time polymerase chain reaction (real-time PCR). A challenge in this field is the fact that real time PCR measures DA copy numbers, whereas legislation will express tolerance limits in terms of mass fractions of PAPs in feedingstuffs. Here we evaluated the impact of different sterilisation temperatures, showing that at high temperatures the DNA copy number was significantly reduced. In addition we showed that measuring at travel levels could lead to a high variation of the measured DNA copy number, characterised by a positively skewed distribution. Various implications of these findings are discussed.

VON HOLST Christoph;  BOIX SANFELIU Ana;  MARIEN Alin;  PRADO Marta; 

2011-11-29

SPRINGER HEIDELBERG

JRC59247

1618-2642,   

www.elsevier.com/locate/foodcont,    https://publications.jrc.ec.europa.eu/repository/handle/JRC59247,   

10.1016/j.foodcont.2011.09.017,