Identification of Processing Adjuvant Trionyx Sinensis Blood and Common Adulterants by AS-PCR

  OBJECTIVE  To establish an allele-specific PCR identification (AS-PCR) method of Trionyx sinensis blood, which can quickly and accurately identify Trionyx sinensis blood from other common animal blood.

  METHODS  Based on the comparison of CO Ⅰ sequences between the Trionyx sinensis and common animals such as donkey, cow, goat, chicken, pig and rabbit, the specific identification primers for Trionyx sinensis blood were designed according to SNP sites, and the PCR reaction system was optimized, and the tolerance, applicability and adulteration sensitivity were investigated and verified.

  RESULTS  When annealing temperature was 62 ℃, the number of cycles was 29, primer dosage was 0.2 μL, DNA template concentration was 100 ng, the bright bands of about 286 bp appeared in the Trionyx sinensis blood samples after PCR amplification and gel electrophoresis with the designed ZHB286 primer, while no bands appeared in other animal blood samples.

  CONCLUSION  AS-PCR identification method has high specificity, can accurately identify the blood of Trionyx sinensis and other common animals, and can be used for the identification of adulterated Trionyx sinensis blood with donkey, cow, goat, chicken, pig and rabbit blood, which has wide application value in the molecular identification of blood medicinal materials.