The study of mitochondrial D-loop region on pangolin product identification

• Main Authors: Chao-An Chen, 陳昭安
• Other Authors: Li-Chin Tsai
• Format: Others
• Language: zh-TW
• Published: 2011
• Online Access: http://ndltd.ncl.edu.tw/handle/ss8hhv

碩士 === 中央警察大學 === 鑑識科學研究所 === 99 === The flesh and scales of pangolin has been one of the important components of traditional Chinese medicine. It incurs hunting and smuggling to pangolin and leads to a sharp drop of its number, which comes to induce its listing on the CITES. The seized scales of pangolin from traditional Chinese medicine are mostly old or sauted, which results in serious decline and fragmentation of DNA. In Taiwan, there is no systemic method to analyze samples and products from pangolin yet. With the samples of different qualities and 5-step strategy, 29 pairs of primers are designed in this study to amplify different fragments in the D-loop region of mitochondrial DNA. In the 122 samples, 15 of them are tissues and blood of Formosan Pangolin. They’re all amplified with full-length sequences and contain 8 haplotypes. The other 107 samples are scales of pangolin. (1 untreated, 22 fried with traditional Chinese medicine, and 84 sauted.) 35 of them gave full-length sequences after analyzing, which can be categorized into 5 subtypes of Formosan Pangolin and 5 subtypes of other pangolins. The incomplete sequences of the remaining 72 scale samples are compared with the GenBank database and few of them are identical to the logged data (mostly unlogged.) For the best efficiency of DNA amplification, some strategies of PCR are established in this study, and they worked successfully on identification of one suspected pangolin powder sample. To sum up, 10 subtypes of Formosan Pangolin and 5 subtypes of other pangolins yielded in this study, and 12 sequences that are unlogged on GenBank occurred after systemic analyzing. It shows that the D-loop region of pangolin has highly polymorphism. The system established in this study has been applied to identify one suspected pangolin powder sample and two specimens successfully. PCR strategies according to the sample of different qualities are developed to get the longest sequences of D-loop region, which is helpful in the identifications of unknown sample and maternal inherence.