以PCR方法與市售免疫套組檢測基因改造大豆   全文下載

Detection of Genetically Modified Soybeans by PCR Method and Immunoassay Kits

以PCR方法與市售免疫套組探討鑑別檢測基因改造大豆之可行性。針對Roundup ReadyTM基因改造大豆（RRS, Monsanto 公司）殖入基因與品種特性基因選定不同引子，進行PCR與ELISA方法檢測。用以鑑別基因改造大豆之引子共四對，分別為35S（針對35S-promoter，源自cauliflower mosaic virus）、NOS（nopaline synthase-terminator, 源自Agrobacterium tumefaciens）、EPSPS（5-enolpyruvylshikimate-3-phosphate synthase，源自A. tumefaciens strain CP4）及LE （品種特性基因lectin）設計檢測。結果顯示，大豆檢體以35S及EPSPS引子檢測時，其最低檢測量均為0.1%（w／w），NOS 則為1%（w／w）；檢體並以LE引子-PCR反應確定均為大豆產品。經以不同免疫檢測套組試驗，條片法較適用於定性之檢測，易於操作、攜帶和判讀；而ELISA方法除可鑑別基因改造大豆外，亦適用於含量在0-2.5%之檢體定量檢測。以市售免疫檢測套組檢測的結果與PCR法檢測的結果相符，均可區分鑑別一般大豆與RRS基因改造大豆。"

The suitability of detecting genetically modified (GM) soybeans by polymerase chain reaction (PCR) and immunoassay kits is determined. Primers specific for inserted genes in the Roundup ReadyTM soybean (RRS, Monsanto company) were applied. Four pairs of primers, namely, 35S (35S-promoter, originated from cauliflower mosaic virus), NOS (nopaline synthase-terminator, derived from Agrobacterium tumefaciens), EPSPS (5-enolpyruvylshikimate-3-phosphate synthase, obtained from A. tumefaciens strain CP4) and LE (endogenous gene lectin) were used to identify the GM soybeans. The detection limit of PCR was 0.1% (w/w) GM soybeans when primers 35S and EPSPS were used, and 1% when primers NOS were used. All soybean samples were evidenced by LE primer-PCR as soybean products. Results of Immunoassays by two kinds of kits, strip and ELISA were matched with PCR’s, and feasibility of quantitation detecting GM soybeans is determined among 0~2.5%. The data further revealed that the PCR method and immunoassay kits can sufficiently differentiate GM soybeans from non-GM products. 以PCR方法與市售免疫套組探討鑑別檢測基因改造大豆之可行性。針對Roundup ReadyTM基因改造大豆（RRS, Monsanto 公司）殖入基因與品種特性基因選定不同引子，進行PCR與ELISA方法檢測。用以鑑別基因改造大豆之引子共四對，分別為35S（針對35S-promoter，源自cauliflower mosaic virus）、NOS（nopaline synthase-terminator, 源自Agrobacterium tumefaciens）、EPSPS（5-enolpyruvylshikimate-3-phosphate synthase，源自A. tumefaciens strain CP4）及LE （品種特性基因lectin）設計檢測。結果顯示，大豆檢體以35S及EPSPS引子檢測時，其最低檢測量均為0.1%（w／w），NOS 則為1%（w／w）；檢體並以LE引子-PCR反應確定均為大豆產品。經以不同免疫檢測套組試驗，條片法較適用於定性之檢測，易於操作、攜帶和判讀；而ELISA方法除可鑑別基因改造大豆外，亦適用於含量在0-2.5%之檢體定量檢測。以市售免疫檢測套組檢測的結果與PCR法檢測的結果相符，均可區分鑑別一般大豆與RRS基因改造大豆。