Development and validation of a TaqMan-based real-time PCR assay for quantifying chicken meat in minced meat products

Abstract

This study describes a novel quantitative real-time PCR assay to quantify chicken content in commercial minced meat products. This method used chicken-specific primer pair and probe targeting the mitochondrial 16S rRNA region to amplify the target without cross-reactivity, achieving a limit of detection (LOD) of 0.1 pg and a limit of quantification (LOQ) of 1 pg for raw chicken meat. For autoclave-treated meat, both the LOD and LOQ were 1 pg. Calibration curves generated using reference mixtures confirmed the method's suitability for quantification. This approach was applied to 24 commercial minced meat products, including meatballs, nuggets, and patties. Intra- and inter-laboratory validations were performed using three types of equipment and two operators, demonstrating high sensitivity and specificity. Therefore, this method provides reliable and accurate quantification of chicken content in commercial minced meat products.