A Real-Time Quantitative PCR Detection Method for Pork, Chicken, Beef, Mutton, and Horseflesh in Foods

TLDR

A rapid real‑time quantitative PCR method was developed to detect trace amounts of pork, chicken, beef, mutton, and horseflesh in foods. Primers and TaqMan MGB probes targeting cytochrome b were designed for species‑specific detection. The assay achieved a limit of quantification of 100 fg/µl mitochondrial DNA in a wheat matrix, with standard‑curve R² values between 0.994 and 0.999, making it suitable for detecting hidden meat mince and verifying food labeling.

Abstract

A rapid real-time quantitative PCR method to detect trace amounts of pork, chicken, beef, mutton, and horseflesh in foods was developed. The primers and TaqMan MGB (minor groove binder) probes were designed on the gene encoding cytochrome b for the specific detection of each species. The limit of quantification of this method was found to be 100 fg/μl of each mitochondrial DNA in 10 ng/μl of the wheat mitochondrial DNA matrix. The calculated R2 values of the standard curves for the five species ranged between 0.994 and 0.999. This method would be particularly useful in the detection of hidden meat mince in processed foods, which would verify food labeling and gain consumers' trust.