Optimisation of Real-Time Polymerase Chain Reaction Methods for Accuracy and Precision

The polymerase chain reaction (PCR), with endpoint determination, is a powerful tool for food analysts in all fields since it allows the specific amplification and detection of as little as a single copy of a particular nucleotide sequence. It therefore has great potential for detection of adulteration and contamination. Recently, real time quantitative PCR (Q-PCR) has been introduced. Q-PCR measures the accumulation of PCR products in real time, thereby overcoming the drawbacks of end point analysis, enabling the quick and easy quantification of nucleic acids and by extrapolation, weight or percentage of analyte. Although the use of Q-PCR is becoming widespread, it should be used with caution and the results should be carefully scrutinised as there are issues with accuracy and precision particularly when looking at processed samples.