Development of a fast duplex real-time PCR assay for simultaneous detection of chicken and pigeon in raw and heat-treated meats

We developed a fast duplex real-time polymerase chain reaction assay based on TaqMan (R) probes for the simultaneous identification of chicken and pigeon meat in under 40 min. The primer sets used for both pigeon and chicken targeted the mitochondrial cytochrome b gene. To obtain simultaneous fluorescent signals, FAM and HEX probes were used to label the pigeon- and chicken-specific probes, respectively. This method specifically amplified chicken and pigeon DNA without any cross-reactivity to 20 animal species. The absolute detection limit was 0.1 pg of DNA extracted from both chicken and pigeon meat. Using DNA extracted from either raw or heat-treated meat, as little as 0.01% pigeon DNA was detectible in chicken DNA, as well as the reverse. This assay might serve as a rapid, sensitive, and specific detection method for the identification of pigeon and chicken meats in both raw- and cooked-meat products. (C) 2017 Elsevier Ltd. All rights reserved.