Direct or DNA Extraction-Free Amplification and Quantification of Foodborne Pathogens
The use of direct nucleic acid amplification of pathogens from food matrices has the potential to reduce time to results over DNA extraction-based approaches as well as traditional culture-based approaches. Here we describe protocols for assay design and experiments for direct amplification of foodborne pathogens in food sample matrices using loop-mediated isothermal amplification (LAMP) and polymerase chain reaction (PCR). The examples provided include the detection Escherichia coli in milk samples and Salmonella in pork meat samples. This protocol includes relevant reagents and methods including obtaining target sequences, assay design, sample processing, and amplification. These methods, though used for specific example matrices, could be applied to many other foodborne pathogens and sample types.
Key wordsDirect amplification Direct loop-mediated isothermal amplification Direct polymerase chain reaction Foodborne pathogens
Support for this research was provided in part by a grant from the National Institute of Environmental Health Sciences, National Institutes of Health grant number P42ES04911-26.
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