赭麴毒素A(ochratoxin A)與微囊藻毒(microcystin-LR)是作物與
飲水中兩種常見到的小分子毒素，食用遭受污染的食品與飲水
會導致人類許多疾病及癌症的生成。由於此類毒素對於人類及
動物體健康有高度的威脅，因此開發各式抗體及建立快速檢測
分析法來分析調查各類食品中赭麴毒素與微囊藻毒的污染是一
重要課題。本計畫主要將建立赭麴毒素A 與微囊藻毒之螢光極
化免疫分析法(fluorescence polarization immunoassay, FPIA)來檢
測分析食品作物中與飲水中此二類毒素污染情形。其原理是利
用抗體與小分子毒素-螢光接合物鍵結飽和時，螢光偏極化值最
大，當加入小分子毒素時，毒素與定量之毒素螢光接合物兩者
競爭固定量的抗體，當外加毒素(或樣品)濃度高時，較少量的
毒素螢光接合物接合到抗體，因此螢光偏極化值變小，此螢光
偏極化數值可由儀器直接測得，而且穩定性極佳。因此首先我
們先合成各赭麴毒素A(ochratoxin A)與微囊藻毒(microcystin)的
不同螢光衍生物，接著將此毒素與螢光衍生物與不同毒素濃度
之標準品與其專一性抗體進行反應，以建立毒素螢光偏極化標
準曲線，針對微囊藻毒各種類似物MCY-LR, YR MCY-YR,
MCY-RR 和nodularin 分別為46, 60, 172,and 45 ng/mL。針對赭
麴毒素A 所建立之螢光極化免疫分析50%抑制濃度(IC 50)大約
在98 ng/mL 左右。目前正利用此方法進行各類樣品分析。
Ochratoxin A and microcystin-LR are two natural carcinogenic
toxins. They are
frequently contaminated in food, feed and water which cause toxic
effects and cancer
in human and animal. Production of antibodies and development of
rapid detection methods for ochratoxin A and microcystin are
important issue in food safety. This study is to establish these
fluorescence polarization immunoassays (FPIA) for ochratoxin A
and microcystin-LR and apply this technology for rapid monitoring
food, feed and water contamination.of this two toxins. FPIA is
based on the increase in polarization of the fluorescence of
fluorescent-labeled toxin (tracer) binding with specific antibody.
When the free toxin is added to compete with fluorescent-labeled
toxin for the limited antibody. The higher free toxin is added, the
lower fluorescence polarization is measured. Using different MCYLR
analogs to test the established FPIA.The IC50 for MCY-LR,
MCY-YR, MCY-RR and nodularin are 46, 60, 172, and 45 ng/mL,
respectively. While the IC 50 for the ochratoxin A of FPIA is 98
ng/mL. Currently, we are woking on sample analysis using these
established FPIA methods.
