鱈屬魚類Real-time PCR快速鑑別檢驗方法之建立   全文下載

Rapid Identification of Cod (Gadus spp.) in Food using Real-time PCR

水產品經切片或加工後辨識不易，易發生業者以低價魚種混充高價魚賺取價差之詐欺事件，為解決水產品容易誤判之現象，本研究著重於鱈屬（Gadus spp.）魚種鑑別檢驗，包括大西洋鱈（Gadus morhua）、太平洋鱈（Gadus macrocephalus）、格陵蘭鱈（Gadus ogac）、黃線狹鱈（Gadus chalcogramma）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120個台灣市售魚種進行測試，確認本方法對鱈屬魚種具有專一性，針對不同濃度的鱈屬魚肉DNA，本方法靈敏度可達1pg DNA，相較於透過比對PCR產物序列的生物條碼鑑種方法，本方法單次反應可檢驗4個鱈屬物種，包含DNA萃取與real-time PCR鑑定過程可於4天內完成，本研究開發之鱈屬快速鑑別檢驗方法，可實際應用於鱈屬魚類產品之快速檢驗，維護民眾權益，為國人消費飲食把關。

Seafood mislabeling related to economic fraud or trade dispute frequently occurs all over the world. The processing step such us slicing of aquatic products may lead to substitutions of species deliberately, which results in increased difficulties of species identification. It is necessary to develop methods that permit the accurate identification or called authentication of the species present in all types of processed fish products. This study aimed at the prevention of fish product mislabelling, a rapid method of identification utilizing real-time PCR was developed for cod (Gadus spp.), including Gadus morhua, Gadus macrocephalus, Gadus ogac and Gadus chalcogramma. The primers and Taqman probes were designed specifically to the gene encoding cytochrome b (Cyt b) as barcode marker. Specificity was confirmed by testing 120 fish species frequently presented in the markets of Taiwan. The detection limit was as low as 1 pg of DNA. These data showed our real-time PCR method provided a rapid, sensitive and reliable detection tool for accurate identification of cod products, allowing the detection of fraudulent and unintentional mislabeling of this species.