- 學位論文
同時檢測魚蝦主要過敏原之側向流體免疫分析法
Simultaneous detection of major allergens in fish and shrimp by lateral flow immunoassay
摘要
近年來食物過敏議題逐漸引起大眾重視，台灣主要過敏性食物為海鮮。魚中小清蛋白β (β-Parvalbumin) 及蝦中原肌球蛋白 (Tropomyosin)是患者食用魚蝦相關食品導致過敏的主要過敏原，食物過敏輕微誘發嘔吐腹瀉、紅腫發癢，重則呼吸困難休克致死，因此發展適合社會大眾之快速定性過敏原的分析方法是必要的。側向流體免疫分析法 (Lateral flow immunoassay, LFIA) 是藉由抗體與抗原專一性結合達到快速定性特定目標抗原的方法，由於傳統側向流體免疫分析法通常只能檢測單一抗原，本研究目的是建立一種奈米金標記之新型側向流體免疫分析法且能夠同時檢測甲殼類原肌球蛋白及魚類小清蛋白β。本研究在競爭型側向流體免疫分析法基礎上；使用針對蝦原肌球蛋白、魚小清蛋白β之二種兔子多株抗體 (Rabbit IgG, pAb) 並結合奈米金標記作為偵測抗體 (Detection antibody) 以檢測樣品中目標過敏原；後使用純化之蝦原肌球蛋白、魚小清蛋白β製作測試線1 (Test line 1)、測試線2 (Test line 2)；使用Goat Anti-Rabbit IgG製作控制線針對偵測抗體進行有效測試，即可實現同時檢測甲殼類原肌球蛋白、魚小清蛋白β之競爭型側向流體免疫分析法。結果在無論何種情況下控制線首先需要顯示以表示條帶免疫系統之有效性；後根據測試線1、測試線2 顯示與否；分別定性樣品是否含有甲殼類原肌球蛋白、魚小清蛋白β。樣品測試部分，在4種甲殼類樣品之結果均能檢測到甲殼類原肌球蛋白。在3種魚類樣品之結果均能檢測到魚小清蛋白β。顯示出本方法能檢測甲殼類原肌球蛋白及魚小清蛋白β且不會與其他物種交叉反應，結果也與先前研究文獻一致，預期本方法能夠在食品及相關醫療產業上達到製程控管、鑑別食品過敏原之貢獻。
並列摘要
In recent years, the issue of food allergy has gradually attracted public attention. The main allergens in Taiwan are seafood. Fish β-parvalbumin and shrimp tropomyosin are the main allergens, caused patients get food allergies by eating shrimp and fish, mild food allergies induce vomiting, diarrhea, redness, swelling and itching, and severe breathing difficulties can lead to shock and death. Therefore, it is necessary to develop detection methods suitable for the public. Lateral flow immunoassay (LFIA) uses specificity of combine of antigen and antibody to determine allergen. Because the traditional LFIA can only detect one antigen, the purpose of this research is to develop a lateral flow immunoassay of goldparticle labeled can detect two allergens at same time. This method simultaneously includes two different species: fish and crustacean, it has a short test time and low technical requirements for operators. Based on the competitive LFIA, uses two rabbit IgG combined with goldparticle label as detection antibody, against crustacean tropomyosin and fish parvalbumin beta. Uses shrimp tropomyosin and fish β-parvalbumin to make test line 1 and test line 2. Uses Goat Anti-Rabbit IgG to make a control line for valid test, then this new type competitive LFIA can achieve detecton of crustacean tropomyosin and fish β-parvalbumin at same time. The result is expected that control line needs appear first under any situation to indicate the effectiveness of the strip immune system; then according to test line 1 and test line 2 are appear or not, to determine sample contains crustacean tropomyosin or fish β-parvalbumin. In the sample testing part, the results of the four crustacean samples were all can dectect crustacean tropomyosin. The results of the three fish samples were all can dectect fish β-parvalbumin. Indicating that the method did not cross-react tropomyosin and β-parvalbumin from other species. The results of this study are consistent with those of previous literature, this method is expected can achieve the contribution of process control and identification of food allergens in the food and related medical industries.
參考文獻
延伸閱讀
- 余旭勝（2019）。食品中蝦過敏原之質譜檢測方法開發。屏科大研發專刊，()，5-5。https://www.airitilibrary.com/Article/Detail?DocID=P20110422001-201912-202105210010-202105210010-5-5-005
- Sung, H. H., & Su, Y. J. (2005). 壬基苯酚和壬基苯酚單乙氧基醇化物對淡水長臂大蝦血球細胞的影響：活存力及免疫反應之體外試驗. 臺灣水產學會刊, 32(2), 101-111. https://doi.org/10.29822/JFST.200506.0002
- Tsai, J. M. (2006). 蝦類白點症病毒結構性蛋白質之鑑別及研究 [doctoral dissertation, National Taiwan University]. Airiti Library. https://doi.org/10.6342/NTU.2006.01290
- Chin, W. C., Huang, C. C., & Wang, J. Y. (2020). Analysis of gut microbiota composition of aquaculture fish and establishment of predominant flora. 屏科大研發專刊（英文版）, (), 3-4. https://www.airitilibrary.com/Article/Detail?DocID=2076880X-202012-202105270009-202105270009-3-4-020
- 周薰修、賀克勤、陳漢恆（1986）。Analysis of Polychlorinated Biphenyls in Fish Shellfish from Taipei County。化學，44(2)，43-53。https://doi.org/10.6623/chem.1986015