Rapid identification of dried suckling deer using molecular authentication-based PCR-GICA and PCR-FQICA

Dried suckling deer is an animal-derived traditional Chinese medicinal material that is vulnerable to adulteration in the marketplace because of its limited availability, relatively high price, and lack of unified quality standards. Therefore, reliable and rapid authentication of dried suckling deer is essential. This research aimed to establish rapid PCR-based colloidal gold immunochromatographic assay (PCR-GICA) and PCR-based fluorescent quantum dot immunochromatographic assay (PCR-FQICA) methods based on molecular authentication technology for the identification of dried suckling deer. The mtDNA ND4L gene of dried suckling deer was selected as the target gene, and dried suckling deer-specific primers were designed to amplify a 46 bp fragment. PCR amplification was performed, and the amplified products were rapidly detected using PCR-GICA and PCR-FQICA, with agarose gel electrophoresis used for verification. The specificity, reproducibility, and sensitivity of the two methods were evaluated. The results showed that the short-fragment primers designed from the mtDNA ND4L gene achieved the best specificity at an annealing temperature of 56 °C. Both PCR-GICA and PCR-FQICA showed positive results for the dried suckling deer reference material and authentic dried suckling deer samples, whereas the confusing product samples and the blank controls showed negative results. Both methods showed good specificity and reproducibility, and the lowest detectable concentration of template DNA was 0.5 pg/μL, which was 10-fold lower than that of agarose gel electrophoresis. In conclusion, the rapid identification methods established in this study are specific, sensitive, rapid, low-cost, and visual, and are suitable for the rapid identification of dried suckling deer.