High‐resolution melting analysis to authenticate deer‐derived materials in processed products in China using a cytochrome oxidase I mini‐barcode

BACKGROUND Deer‐derived materials (antler, venison, fetus, penis, bone, tail, and others) are some of the most valuable traditional animal‐based medicinal and food materials in China. In production, processing, and trade, the quality of deer products varies. The market is confusing, and counterfeit and shoddy products are common. There is an urgent need to establish an accurate identification method. RESULTS Two pairs of primers suitable for identifying deer‐derived medicinal materials were obtained by screening the cytochrome oxidase I (COI) sequences of 18 species from nine genera of the deer family. The two primers were used to identify the species and adulteration of 22 batches of commercially available deer‐derived products with a mini‐barcode combining high‐resolution melting (HRM) technology and methodical investigation. Deer‐derived materials (sika and red deer) were correctly identified by species using varying DNA amounts (1 to 500 ng). The two pairs of primers COI‐1FR and COI‐2FR yielded melting temperatures (Tm) of 80.55 to 81.00 °C and 82.00 to 82.50 °C for sika deer, and 81.00 to 82.00 °C and 81.40 to 82.00 °C for red deer. Twenty‐two batches of commercially available samples were analyzed by HRM analysis and conventional amplification sequencing, and it was found that the species samples had an error rate of species labeling of 31.8%. Four batches of samples were identified as mixed (adulterated) in the HRM analysis. CONCLUSION The combination of DNA mini‐barcode with HRM analysis facilitated the accurate identification of species of deer‐derived materials, especially the identification of samples in an adulterated mixed state. © 2024 Society of Chemical Industry.