以增幅細胞色素b基因片段鑑定家犬、家貓及美洲巨水鼠來源肉品

Abstract

在許多東亞社會中，因為肉用、毛皮或者為其他需要，許多狗貓被不人道地任意屠宰。隨著台灣動物保護法的實施，肉品之物種鑑定技術便成為行政機關對不法業者處罰時的重要依據。此外，有一些有害外來物種如美洲巨水鼠(Myocastor coypus)遠赴重洋地從原棲地經由走私入侵台灣，並且可能逐漸對台灣溼地生態系統造成傷害。對於此類有害外來種生物的鑑定結果，亦為其非法進口、飼養及販賣之違法行為之重大證據。本論文係以增幅粒線體DNA上之cytochrome b基因片段來作為鑑定方法，對一些品質不良的檢體，例如腐壞或者過度烹煮的肉或者毛皮仍然適用，在整個鑑定過程中，每個檢體只需要萃取出15 ng的DNA便足以辨識。本鑑定法係利用三對專一性的引子將各標的物種cytochrome b基因片段以是否分別增幅出252、292及297 bp大小的片段來判斷為標的物種與否。本實驗已將所得PCR產物的定序結果及與已在EMBL DNA資料庫上發表的序列進行比對，結果均確實無誤，以確保各鑑定結果的正確性。

Many dogs and cats are slaughtered in East-Asia because of the needs in their meat for food or other purposes. Identification of the species is required to determine that a crime against Animal Protection Law in Taiwan has been committed. Otherwise, some harmful species like nutria (Myocastor coypus) has invaded Taiwan far from the place they originally inhabit, and may damage the ecological system of wetland in Taiwan gradually. This thesis details a good DNA technique using part of the cytochrome b gene on the mitochondrial genome that can work on poor-quality samples such as decayed or over-cooked meat. An appropriate deoxyribonucleic acid (DNA) extraction technique is required to obtain at least 15 ng of DNA. Amplification of part of the cytochrome b gene using specific primers produces a fragment of 252/292/297 bp in size. Direct sequencing of the PCR products allows confirmation of the DNA sequences to those already described on the GENbank-EMBL DNA database to ensure the results of identification.