利用短分散重複序列（SINE）偵測深度加工龜甲（龜甲膠）中之烏龜 DNA   全文下載

Tortoise DNA Detection from Highly Processed Tortoise Shell Using SINE Element

龜甲膠（Colla carapax et plastrum testudinis）是一種由龜科動物──烏龜的背甲或腹甲經水煎煮、濃縮製成的固體膠，是一種傳統名貴中藥。目前中藥市場上存在很多假冒產品其製備原料並非龜甲，而是其他動物的組織或雜皮，大大地損害了消費者的權益。龜甲膠是一種深度加工型產品，DNA破壞降解嚴重，含量極少，片段極短。基於此，本研究選取拷貝數極高的Tortoise PolIII/SINE序列，建立了用於龜源性成分檢測的DNA分子鑒定方法。可將龜基因組與其他動物基因組包括驢、馬、牛、豬進行區分。本方法能夠成功檢測僅含 0.1% 龜甲膠的混合膠樣，靈敏度極高。此外，還比較了5種PCR聚合?應用於本方法之檢測情形，發現差異並不明顯。最後，將本方法實際用於市售6種龜甲膠產品之鑒別，其中有一樣品之檢測結果呈陰性，說明該產品極有可能是假冒偽劣之產品或者其龜甲膠含量低於0.1%。

Colla carapax et plastrum testudinis is one of the most valuable traditional Chinese medicine referring to highly processed tortoise shell glue which contains trace amount of severely degraded DNA. In this work, a simple and fast methodology was first developed for the identification of highly processed medicine from substitutions which were labeled fraudulently as tortoise shell glue but derived from tissues of other animals such as horse, cattle or pig. The method is based on polymerase chain reaction (PCR) amplification of tortoise-specific Pol III/SINE element and allows the detection of tortoise DNA in glue mixtures, which contain only 0.1% colla carapax et plastrum testudinis. A comparison of five various polymerases for the assay was also conducted, based on their performances in PCR. Finally, the method was applied for the detection of tortoise DNA from six commercial tortoise shell glue products; one of them was tested negative, indicating none or less than 0.1% tortoise shell in there.