| Summary: | 碩士 === 中山醫學大學 === 生物醫學科學學系碩士班 === 99 === 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.
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