A型肉毒毒素胶体金免疫层析法检测试纸条的研制

Development of gold-immuochromatography assay test paper for botulinum toxin type A

目的 :研制一种敏感、特异、快速并适用于非专业人员的胶体金免疫层析法 (immuno_chromatographicassay ,ICA)试纸技术 ,用于检测食物、患者呕吐物或粪便以及血清和土壤中的肉毒毒素 (botulinumtoxin)。方法 :采用柠檬酸盐还原法制备胶体金颗粒、标记A型肉毒毒素的多克隆抗体 ,制成免疫层析检测试纸条。待测样中的A型肉毒毒素与试纸条上金标记抗体结合后沿着硝酸纤维膜移动 ,并与膜上的抗体结合形成肉眼可视红色带。结果 :肉毒毒素ICA试纸条可以特异性检测A、B和E型肉毒毒素。检测中国兰州生物制品研究所注射用A型肉毒毒素标准品 ,灵敏度可达 2U ml。与金黄色葡萄球菌肠毒素等抗原无交叉反应。结论 :A型肉毒毒素ICA试纸条具有特异性强、灵敏度高、简便快速的特点 ,适用于专业和非专业人士 ,实用性较强。

Objective:To develop sensitive,specific, rapid, simple and selfperforming gold immunochromatography assay (ICA) test paper for detection of botulinum toxin in soil and food, puke, egesta and serum of patients.Methods: Colloidal gold was coupled with polyclonal anti-botulinum toxin antibody to prepare an ICA test paper. The gold-anti-botilinum toxin conjugate reacts with botulinum toxin in samples and migrates up the nitrocellurose strip on which anti-botulinum toxin antibody was immobilized, thus producing the detection reaction.Results:Botulinum toxin types A,B,E could be detected by ICA test paper,while the enterotoxin could not. The sensitivity for standard botulinum toxin from Lanzhou Institute of Biological Products was 2?U/ml.Conclusion:ICA test paper is a sensitive,specific,simple and rapid assay for detecting botulinum toxin.