扁鱈(大比目魚)real-time PCR快速鑑別檢驗方法之建立   全文下載

Rapid Identification of Halibut (Hippoglossus spp. And Reinhardtius spp.) Using Real-time PCR

水產品經切片或加工後外觀特徵消失，易發生業者以攙偽、假冒、標示不實等方式牟利，以較低價魚種混充高價魚賺取價差，這類詐欺事件在全球屢見不鮮，本研究針對扁鱈（俗稱大比目魚）魚種鑑別檢驗，扁鱈物種為鰈形目之庸鰈（Hippoglossus hippoglossus）、狹鱗庸鰈（Hippoglossus stenolepis）及馬舌鰈（Reinhardtius hippoglossoides），對於上述三個物種開發分子生物快速檢驗方法。以魚類粒線體cytochrome b基因經過多重序列比對，建立專一性引子對及探針，並以即時聚合酶連鎖反應（real-time PCR）進行特異性及靈敏度測試，與125個台灣市售魚種進行測試，確認本方法對扁鱈魚種具有專一性，對於不同濃度的扁鱈魚肉DNA，本方法靈敏度可達10 pg/μL，本研究開發之扁鱈快速鑑別檢驗方法，可實際應用於扁鱈（大比目魚）相關水產品快速檢驗，保護國人消費飲食權益。

Seafood mislabeling related to economic fraud or trade dispute frequently occurs all over the world. Processing and slicing which results in increased difficulties of species identification may lead to mislabeling and substitutions of species deliberately. This study aimed at the prevention of fish product mislabeling, and a rapid method of identification utilizing real-time PCR was developed for halibut (Hippoglossus spp. and Reinhardtius spp.), including Hippoglossus hippoglossus, Hippoglossus stenolepis, and Reinhardtius hippoglossoides. The primers and Taqman probes were designed specifically to the gene encoding cytochrome b (Cyt b). Specificity was confirmed by testing 125 fish species frequently found in the markets of Taiwan. The detection limit was as low as 10 pg/μL of DNA. These data showed the real-time PCR method provided a rapid, sensitive and reliable detection tool for accurate identification of halibut products, allowing the detection of fraudulent and unintentional mislabeling of these species.