Development and Evaluation of A Dog-derived DNA Test Kit

To develop a DNA detection kit for rapid identification and make a methodological evaluation of the kit of dog-derived components in meat products,the genomic DNA of dog meat and adulterated meat(fox meat,clam meat)was extracted by modified SDS-PK method. The mitochondrial cytochrome B gene with species specificity was selected as the target gene for identification. Primer5.0 was used to design specific primers. Dog-derived DNA detection methods,assembly of dog-derived DNA detection kits,and specificity,stability and repeatability were evaluated. The results showed that the dog,fox and mink-derived ingredients in the meat products could be effectively detected by the kit,and DNA was efficiently extracted by the kit. Genomic DNA with a concentration higher than 100 ng/μL could be extracted from only 0.05 g of meat,OD_260/280 was between 1.80~2.00,and the minimum detection limit was 10^-4 ng/μL. All detection steps could be completed within 4.5 h. The advantages of simple operation and high specificity were provided by the kit. The reagents used in the kit were safe and non-toxic. It was still effective after repeated after freeze-thaw cycles for 20 times and it could be stored at-20 ℃ for one year.12 samples were randomly selected for testing. There were 3 samples with fox and mink meat posing as dog meat. The adulteration rate was as high as 25%,which was consistent with the test results of Changchun food and drug inspection center. The dog DNA detection kit had high sensitivity and specificity,accurate and stable detection results,good repeatability,and was suitable for rapid qualitative detection of dog meat.