有機磷農藥二氯松快速檢測試劑之開發

DEVELOPMENT OF RAPID TEST FOR ORGANOPHOSPHORUS PESTICIDES-DICHLORVOS

林秋月(Chiu-Yuen Lin)

指導教授 ： 王鐘毅

大同大學/工程學院/生物工程學系所/碩士(2010年)

摘要

目前現有藥物殘留檢測技術中，以免疫層析最具專一性、快速及低成本等優點。所以本文將以此技術作為研發平台，結合目前已知有機磷農藥為乙醯膽鹼酯酶( Acetylcholinesterase；AchE )之抑制物，且乙醯膽鹼( Acetylcholine；Ach )又為AchE之受質關係，並運用後兩者作為開發檢驗試劑之測試片原料藥劑。就測試片上劑藥之安排與規劃以兔抗AchE抗體作為控制線(Control line；C線)原料，Ach作為測試線（Test line；T線）原料，而AchE聯結膠體金製成複合體作為試劑偵測判讀之訊號物，最後再設計一連串的實驗確認其可行性。
本文開發檢驗試劑為競爭型之試劑，其檢測原理為檢測之目標物與測試片的T線藥劑互相競爭捕捉AchE-膠體金複合體，所以當檢測目標物濃度低於檢驗試劑閾值時，該試劑片上的T線藥劑即能捕獲大部份的複合體，而產生非常清楚的T線。反之，若目標物濃度高於檢驗試劑閾值時，T線的呈色就愈淡或看不見。
總結本實驗以有機磷農藥二氯松作為代表性之測試，在已知溶液中進行實驗：陽性實驗組為二氯松標準品配製在20%甲醇溶劑，濃度為50mg/mL，而陰性對照組則以20%甲醇溶劑，將兩組各別與AchE-膠體金複合體進行反應後，藉由測試片組件毛細管現象，帶動複合體流經測試片C/T線反應區，結果在陰性對照組確實可產生明顯的T線，而陽性實驗組的T線明顯的比對照組呈色淡。故評估此雛型具備檢測農藥殘留之能力。

關鍵字

乙醯膽鹼酯酶 ； 乙醯膽鹼

並列摘要

Among the residual pesticides analysis technologies, immunochromatography strip-test has the benefits of high specificity, rapid detection and low cost.Organophosphorus pesticides are the inhibitor of acetylcholinesterase. Organophosphorus are widely used in agricultural planting. Because this group of pesticides and their secondary metabolites are classified as carcinogens, they are prohibited or restricted to use in many countries. However, their effectiveness and convenient to use make them still been used in some area, which resulted in contamination of agricultural commodities. This study developed a test strip to detect residual dichlorvos in agricultural product. The test strip included acetylcholinesterase and anti-acetylcholine antibody. Acetylcholinesterase was coupled with colloid gold. Anti-acetylcholine antibody was immobilized and used as control line (C-line). Acetylcholine was used in test line (T-line).Dichlorvos appeares in the sample will inhibit acetylcholinesterase and bind to colloid gold complex, which makes the T-line disappear. The results showed that 50 mg/mL dichlorvos can be detected with the test strip.

並列關鍵字

Acetylcholinesterase ； Acetylcholine

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