小分子毒素如藻類毒素及黴菌毒素，在自然界中分佈十分廣泛，易遭人類或動物誤食而對健康造成危害，本研究以黃麴毒素B1 (Aflatoxin B1；AFB1) 與軟海綿酸 (Okadaic acid；OA)為主題，希望分別建立一套能有效檢測這兩類小分子毒素的檢測方式。黃麴毒素B1為一個廣泛汙染飼料和食物的的黴菌毒素，研究指出AFB1具有很強的致肝癌性，國際癌症研究中心 (International Agency for Research on Cancer, IARC) 將其歸類為group 1 ，是對人類確定的致癌物。為了能快速的在食品中檢測到AFB1的含量，我們製備了AFB1的多株抗體，並建立敏感性高的直接競爭型酵素連結免疫吸附分析法 (Competitive direct enzyme-linked immunosorbent assay ；cdELISA)，以及以奈米金粒子為標記物的快速免疫層析試紙分析法。在cdELISA中，抑制50％的AFB1-HRP與抗體結合所需AFB1的濃度 (IC50)為0.156 ng/ml，AFB1專一性的快速免疫層析試紙分析法偵測AFB1的敏感性為2 ng/ml。由檢測食品樣品中AFB1的含量的結果可看出ELISA與免疫層析試紙分析法的結果有很好的一致性，而且免疫層析試紙結果可直接目測並在十分鐘內快速完成。本實驗建立AFB1的cdELISA和免疫層析試紙分析法可用來快速簡便的檢測食品樣品中的AFB1含量。
軟海綿酸為一種常見於貝類中的藻類毒素，人類誤食被OA汙染之貝類後，會有下痢、噁心和嘔吐等症狀，一般被稱為diarrhetic shellfish poisoning (DSP) 症狀。為了發展更快速且敏感的方法來檢測OA，我們製備了OA的多株抗體，以建立敏感性高的cdELISA，以及以奈米金粒子為標記物的快速免疫層析試紙分析法。在cdELISA中，抑制50％的OA-HRP與抗體結合所需OA的濃度 (IC50)為0.94 ng/ml，OA專一性的快速免疫層析試紙分析法偵測OA的敏感性為1 ng/ml。我們分別成功的製備出這兩種小分子毒素的多株抗體，而且開發的cdELISA及免疫層析試紙分析法能快速且有效的檢測出小分子毒素，以避免大眾遭受到這些小分子毒素的危害。
Mycotoxins and phycotoxins are low-molecular-weight nonimmunogenic toxins, which are frequently found in food and feed, and are capable of causing disease and death in humans and other animals. This study focused on Aflatoxin B1 (AFB1) and Okadaic acid (OA), and developed an efficient method to detect the toxin. Aflatoxin B1 is a widespread mycotoxin contaminating food and feed, and found in animal studies to be strong hepatotoxins and potent carcinogens. The International Agency for Research on Cancer (IARC) has classified AFB1 as a class 1 human carcinogen. The AFB1-specific polyclonal antibody was used for developed a sensitive direct competitive enzyme-linked immunosorbent assay (cdELISA), and a gold-nanoparticle based rapid immunochromatographic strip. The concentration causing 50% inhibition (IC50) of binding AFB1-HRP to the antibodies in the cdELISA were found to be 0.156 ng/ml. The detection limit of immunochromatographic strip was 2 ng/ml for AFB1. The results of immunochromatographic strip analysis of AFB1 in food samples were in good agreement with those obtained from cdELISA. Furthermore, the results can be observed directly by the naked eyes, and it can be completed in 10 minutes. These cdELISA and immunochromatographic strip analysis of AFB1 are suitable for the simple and rapid screening of AFB1 without sample cleanup.
Okadaic acid is a phycotoxin, which is easily contaminating various species of shellfish. When ingested it by humans, many gastrointestinal symptoms, involving diarrhea, nausea and vomiting may occur in the first few hours after consumption, also named diarrhetic shellfish poisoning (DSP). The OA-specific polyclonal antibody was produced and for developed a sensitive direct competitive enzyme-linked immunosorbent assay (cdELISA), and a based gold-nanoparticle rapid immunochromatographic strip. The concentration causing 50% inhibition (IC50) of binding OA-HRP to the antibodies in the cdELISA were found to be 0.94 ng/ml. The detection limit of immunochromatographic strip was 1 ng/ml for OA. We have developed polyclonal antibodies against AFB1 and OA, respectively, and the cdELISA and gold-nanoparticle based rapid immunochromatographic strip can be applied to rapid and efficient toxin detection.
