Design of duplex real-time PCR for the detection of chicken meat in mixed meat products

Laboratory PCR techniques used to control the quality of meat products typically detect mitochondrial DNA sequences (mtDNA). The multiplicity of mtDNA copies in the cell provides such PCR techniques a high sensitivity. The high sensitivity of PCR detecting chicken mtDNA can lead to a positive result when applied to analyze mixed meat products containing chicken eggs. Many meat products, particularly some cooked sausages, do not contain chicken meat, while their recipes include chicken eggs or melange. In such cases, PCR detecting mtDNA cannot distinguish correctly the recipe ingredients from the food fraud with cheaper chicken meat. In this work, we carried out a design of realtime PCR which uses nuclear sequences as a target. A separate task was to select primers for PCR for internal control, for which we selected a nuclear sequence conserved for animals and birds as a target. Here we show that using a unique nuclear sequence as a target sequence allows getting rid of positive results in PCR, when the template is DNA, isolated from chicken eggs or mélange. The designed real-time PCR can be used in protocols applied to detect the falsification of the mixed meat products by chicken meat.

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