Evaluation of quantitative molecular biology methods
Abstract:
To evaluate the applicability of selected modern molecular biology methods to reliably detect and quantify meat species around the 1% (w/w) level for enforcement action with a focus on processed food materials. Three techniques were assessed: real-time PCR, droplet digital PCR and a label-free mass-spectrometry (MS) approach. Both qPCR and ddPCR demonstrated good analytical performance at the 1% (w/w) adulteration level for enforcement action Across all adulterant levels investigated, ddPCR generally showed tighter precision estimates, particularly at the 0.1% (w/w) levels and with the highly processed canned meat sample.There were signficant technical issues with MS when applied to processed food. Whilst light chain derived peptides were detected in all unprocessed sample types, no species specific myosin light chain peptides were identified in the processed food materials
Date:
2016
Keywords:
PCR, ddPCR, qPCR, peptides, species
Funding Body:
UK Department of Environment, Food and Rural Affairs
