Abstract
Adding undeclared species in meat-based food is an illicit practice with possible commercial, ethical and consumer health consequences. DNA-based quantitative methods are the most promising ones and their performance can be deeply affected by the quality of the input DNA. In this study, we performed a comprehensive evaluation of the DNA obtained using four common methodologies. DNA was isolated from three experimental food matrices (minced meat, ravioli filling and ragout) prepared with different percentages of beef and pork meat. Hundred and eighty DNA preps, including all matrix/percentage/kit combinations, were obtained and submitted to downstream quality assessment. Mean values and their statistical significance, obtained by parameter analysis and pairwise comparison, were used to calculate a DNA Quality Score (DQS), integrating quality parameters. DQS identified the salting-out based protocol as the best performing method for all the considered food matrices (p < 0.05); in DNA purification from ragout, the Cetyl-trimethylammonium bromide (CTAB) reagent-based protocol showed comparable results to those obtained by the salting-out method (p < 0.05). An optimal DNA purification is the first step for accurate species quantification in complex meat food.
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