Application of multiplex real-time PCR to commercial
products After validation, the multiplex real-time PCR
assay method was applied to commercial deer products,
including blood, meat, and antler acquired from ecommerce
websites or supermarkets in China. DNA was
extracted from samples of each product and analyzed using
the multiplex real-time PCR method to evaluate the
labeling of these products. Reliable results were obtained
from each DNA sample with a distinctive melting curve
profile (Fig. 5) and a Ct value below 30 (Table 3). Melting
curve analysis demonstrated that, in a total of 27 samples,
18 were from deer and the rest were from common
domestic animals (cattle 4; pig 2; horse/donkey 3). Positive
results were obtained for all samples detected, so false
negatives were not a concern. In one third of the products
analyzed, the name of the species displayed on the label
was not in agreement with the species identified using the
PCR method. Some common domestic animals, which are
less valuable than deer, were used as substitutions for deer
products (Table 3). Not only were deer products authenticated,
but the substituted species were discriminated using the
developed method. The species popularly used as a
substitute for deer were identified and the combined primer
set were validated as appropriate for the detection. These results demonstrate that the developed PCR method can be
effectively used in species identification of highly
processed commercial products