Development Tortoise shell PCR Detection Kit and Evaluation Parameter of the Kit

Tortoise shell is the dry breastplate and carapace of the testudinidae animal turtle (Chinemys reevesii (Gray)). Traditional identification methods can't meet the requirements of quality control of Tortoise shell market. We developed a detection kit for Tortoise shell DNA and to investigate the kit's index including specificity, sensitivity, repeatability and stability. The DNA detection kit was assembled by optimized salting out reagent and multiplex PCR reagent. The mtDNA of Tortoise shell was extracted by the kit, the multiplex PCR technique was carried out to authenticate samples of Tortoise shell. The value of mtDNA extracted by the kit was 1.80±0.05.The result of multiplex PCR indicated that two distinct bands at 335bp and 410bp were shown on the agarose gel electrophoresis in genuine Tortoise shell , but there was no band in fake samples. The specificity of the kit was 100%. The detection limit of the kit was 0.025g of each sample. The kit was effective after being frozen-thawed for 20 times, repeatability test indicated the same result through three times. The Tortoise shell DNA detection kit has properties with high specificity, good sensitivity and stability. So it is suitable for the rapid and accurate detection of Tortoise shell.