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

•Fast duplex real-time PCR was developed for the detection of chicken and pigeon.•The detection limits were 0.01% for pigeon or chicken in meat mixtures.•The method simultaneously identified the two meats in under 40 min.•The applicability of this assay was tested using raw and heat-treated meat samples.

We developed a fast duplex real-time polymerase chain reaction assay based on TaqMan® 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.