A novel triplex real-time PCR method for the simultaneous authentication of meats and antlers from sika deer (Cervus nippon) and red deer (Cervus elaphus)

Deer products have been used as a traditional Chinese medicine for over 2000 years. Due to the high price, the meats and antlers are often adulterated with low-value materials. The present study developed a triplex real-time PCR assay to simultaneously identify the DNA from sika deer and red deer in deer products. The triplex real-time PCR approach showed high specificity for 16 animal species, and the method simultaneously amplified an endogenous control to eliminate false-negative results. The absolute limits of detection for raw meat, meat product, and antlers were 0.05–0.025 pg, 0.05–0.025 pg, and 0.1–0.025 pg, respectively, for sika deer and 0.05–0.025 pg, 0.5–0.25 pg, and 2.5–0.25 pg, respectively, for red deer. This approach could determine 0.1% (w/w) adulteration. In conclusion, the established triplex real-time PCR assay displayed high specificity and sensitivity and can be used for authentic animal products from sika deer and red deer.