淡水小龙虾过敏原胶体金快速检测试纸条的研制

Development of colloidal gold strip for rapid detection of the allergen in freshwater crayfish

目的研制虾过敏原胶体金快速检测试纸条。方法利用克隆表达的具有变应原活性的重组淡水小龙虾原肌球蛋白制备虾过敏原单抗和多抗,用胶体金偶联虾过敏原单抗;将过敏原多抗和羊抗鼠IgG分别包被在硝酸纤维膜上,分别作为检测线和质控线,依次将样品垫、胶体金垫、硝酸纤维素膜和吸水纸组装切割成胶体金免疫层析快速检测试纸条。结果使用研制的试纸条检测淡水小龙虾、日本沼虾、虾蛄、河鲫鱼、带鱼、猪肉和牛肉等样品提取液,只有淡水小龙虾出现阳性条带,其余样品均为阴性,无明显交叉反应。此试纸条的检测灵敏度也很高,达到100 ng/ml。结论该试纸条重复性好、操作简单、灵敏度高、特异性强,能快速检测食品中的淡水虾过敏原,具有很重要的应用推广价值。

Objective To develop a colloidal gold strip for rapid detection of allergen in freshwater crayfish. Methods Colloi- dal gold marked with monoclonal antibody and polyclonal antibody against tropomyosin of red swamp crayfish and goat anti - mouse IgG were sprayed onto the nitrocellulose membrane as detection line and control line. Sample pad, gold jet - pad, nitro- cellulose membrane and absorbent paper were assembled and cut into test strips respectively. Results The specificity results showed that only sample of freshwater crayfish appeared the red positive sediment line while other samples were negative. The limit of detection for the strips which could be read by naked eyes was 100 ng/ml. Conclusion The developed test strip showed good repeatablity, easy operation, high sensitivity, strong specificity and rapidity in detection of allergen in freashwater crayfresh, worhty of promotion and application.