Sample Preparation for Multiplex PCR Assays for Food and Agriculture Applications

Abstract

Foodborne pathogens and spoilage microorganisms influence the safety and quality of food. Polymerase chain reaction (PCR)‐based technologies in food diagnostics have become a promising alternative to conventional culturing approaches due to their rapidity, excellent analytical sensitivity and selectivity, and potential for quantification. Although molecular approaches can be extremely effective with pure microbial cultures, the sensitivity can be reduced radically when they are applied directly to food samples, owing to the complexity of the matrix and the presence of PCR‐inhibitory components. In addition, the contamination level of microbial pathogens in food samples is usually very low, making their detection a challenge. In this book chapter, we are presenting various methods that are used to facilitate PCR detection from food samples, including optimization of the DNA amplification conditions by the use of amplification facilitators and sample preparation methods that will either separate the microbial cells from the PCR inhibitors and/or concentrate the microbial cells to detectable concentrations.