利用短分散重複序列（SINE）偵測深度加工阿膠中之驢 DNA   全文下載

Authentication of Equine DNA from Highly Processed Donkey-Hide Glue (Colla Corii Asini) Using SINE Element

阿膠由驢皮經煎煮、濃縮熬得，屬於深度加工型中藥，dna含量極少，片段極短，為了特異性鑑別出阿膠中驢源性成分以及阿膠中可能摻雜的馬源性成分、牛源性成分以及豬源性成分，本文建立了一系列基於高度重複Sine序列的PCR鑑定方法。首先，利用馬科動物Sine序列eRe-1以及馬特異性衛星序列實現了驢源性成分與馬源性成分的區分，分別能夠檢測到含有0.1% 阿膠，1%馬皮膠的混合膠樣。接著利用牛特異性重複序列1.711B bovine repeat以及豬科動物Sine序列PRe-1特異性鑑別了牛源性成分和豬源性成分，分別能夠檢測到含有0.1% 黃明膠和0.1%豬皮膠的混合膠樣。總之，該方法靈敏，特異性好，操作簡單，能夠用於中藥市場上阿膠產品的真偽鑑別，為膠類中藥質量標準的建立提供了新的思路和方向。

In order to detect the DNA from highly processed medicinal donkey-hide glue Colla Corii Asini from adulterants blended or totally substituted with horse, cattle, and pig tissues, various species-specific PCR assays based on highly repetitive SINE elements were established. For horse and donkey tissues, a two-step PCR assay was adopted, including an initial detection of equine DNA based on the equine SINE ERE-1 and a further detection of horse DNA based on the horse specific satellite. The assay can detect the equine DNA and the horse DNA from binary solid glue mixtures spiked with 0.1% Colla Corii Asini and 1% horse-hide glue respectively. In addition, for bovine and porcine detection, the primer pairs were designed upon 1.711B bovine repeat and PRE-1 respectively. The developed methods can detect bovine and porcine DNA from binary solid glue mixtures spiked with 0.1% cattle-hide glue and 0.1% pig-hide glue respectively. The result shows that our developed PCR assays were useful, convenient and sensitive for Colla Corii Asini identification.