橘黴素酵素免疫分析套組及快篩免疫檢測試紙開發確效及試量產

中山醫學大學機構典藏 CSMUIR > 醫學科技學院 > 生物醫學科學學系暨碩士班 > 研究計劃 > Item 310902500/17988

Please use this identifier to cite or link to this item: https://ir.csmu.edu.tw:8080/ir/handle/310902500/17988

Title:	橘黴素酵素免疫分析套組及快篩免疫檢測試紙開發確效及試量產 Development and pilot production of ELISA kit and immunochromatographic strip for citrinin
Authors:	余豐益謝冠輝
Contributors:	中山醫學大學生物醫學科學學系
Keywords:	橘黴素；酵素免疫分析法；抗體；免疫層析試紙 citrinin；ELISA；antibody
Date:	2017
Issue Date:	2017-07-13T09:32:41Z (UTC)
Abstract:	橘黴素(Citrinin，縮寫CTN，分子量250.25)是由黃麴菌(Aspergillus),青黴菌(Penicillium)和紅麴菌(Monascus)等菌株所產生的二級代謝物，常在玉米、小麥、稻米、裸麥和堅果類等穀物及糧食中被發現。由於紅麴食品含有抗血壓及降膽固醇等有效成分而成為近年來熱門的健康食品，食用遭受橘黴素素污染的食品與飼料會導致人類或動物許多疾病及癌症的生成。國際癌症協會已將橘黴素歸類為C族顯示其對動物是一可能的致癌物，因此開發各式快速檢測分析法來分析調查各類食品中與飼料中的橘黴素的污染是一重要課題。目前本實驗室針對橘黴素已經生產獲得具有專一性極佳之多株抗體並持續努力開發其單株抗體，利用多株抗體已經建立非直接與直接競爭型酵素免疫分析法來分析調查各類食品與紅麴食品的橘黴素。近年來，由於橘黴素汙染食品與紅麴食品的問題日趨嚴重，購買使用橘黴素免疫分析套組日漸增加，從國外進口來的橘黴素酵素免疫分析套組又非常昂貴，因此本計畫將以自行生產之橘黴素專一性抗體進行橘黴素酵素免疫分析套組之開發，將2小時可以完成的檢測分析時間，藉由抗體首先吸附在ELISA plate 上，再調整適當的毒素與酵素接合物的濃度，縮短至30分鐘內來完成檢測結果。此外酵素免疫分析法(ELISA)雖比化學分析法相具有快速而且高靈敏度等優點，但是仍受限於反應所需時間與繁複的反應步驟、需要一些儀器設備與訓練有素之人員材料等。近年來快速,簡單而且靈敏的快速篩檢技術已成為主流，其中奈米科技的進展使得應用奈米金粒子於生物檢測及分析上已快速發展，將奈米金粒子與抗體相結合形成探針來開發出免疫層析試紙(immunochromatographic strip)。我們將以橘黴素之抗體進行奈米快速免疫檢測試紙開發，將紅色奈米金粒子與抗體結合形成抗體奈米金探針，藉由免疫層析試紙的裝置使得檢測耗時1~2小時之ELISA免疫分析縮短至十分鐘內完成，不須要訓練有素人員及任何儀器即可直接目視結果，適合推廣至一般大眾使用。本計畫擬將研究成果與尖端生技公司產學合作進行試量產，期能建立台灣生技公司橘黴素免疫分析套組自有品牌，除了可以提昇產業檢驗試劑生產技術供國內業界使用，並可外銷至其他國家，對於檢驗試劑生技產業將有相當大的助益。 Citrinin (CTN) is a mycotoxin which is produced by Asperillus, Penicillium and Monascus strains. CTN is commonly found in corn, wheat, and Monascus health food products and is known to cause various toxic effects on human and livestock. Due to the red yeast rice food contains blood pressure- and cholesterol-reducing active ingredients and become popular health food in recent years. Consumption of food and feed contaminated with citrinin can cause human or animal diseases and even cancer generation. The International Agency for Research on Cancer (IARC) has classified citrinin as a group C as a possible carcinogen for animals and therefore developed a rapid assay to detect citrinin in food and feed is an important issue. Recently, our laboratory have developed specific polyclonal antibodies for citrinin and continued efforts to develop its monoclonal antibody. By using this polyclonal antibodies, both indirect and direct competitive enzyme-linked immunosorbent assay (ELISA) for citrinin have been established to analyze the various types of food and red yeast rice products.Owing to the citrinin contaminated in food and red yeast rice food is becoming more and more serious, the purchase of the citrinin ELISA kit is increasing, the imported ELISA kit from abroad is also very expensive. Therefore, this project will develop an ELISA kit for citrinin using our specific polyclonal. The antibodies are first adsorbed onto the ELISA plate and by adjusting the toxin-enzyme conjugates concentration in the ELISA will allow the assay to complete from 2 hour to 30 min. In addition, although ELISA is faster than the chemical method for citrinin analysis, but still limits by the reaction time and complex reaction steps, the need for some equipment and well-trained personnel. In recent years, rapid, simple and sensitive rapid screening technology has become the main stream, The application of nanotechnology in biological detecting is on the cutting edge of trends. The conjugation of gold nonoparticles and antibodies as a probe is use to develop an immunochromatographic strip assay. Using our specific antibody, we will develop immunochromatographic strips (immunostrip) for CTN. The immunochromatographic test strip device can be completed within 10 min, without the need for trained personnel and any equipment can be directly visual results. It is suitable for general public use. Above achievement will cooperate with Taiwan Advance Bio-Pharmaceutical Inc. for pilot production. Our goal is to establish Taiwan own brand-name immunoassay reagents for citrinin not only domestic use not also can be exported to other countries. This project will benefit for our biotechnology industry and popular user.
URI:	https://ir.csmu.edu.tw:8080/ir/handle/310902500/17988
Appears in Collections:	[生物醫學科學學系暨碩士班] 研究計劃

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