石杉碱甲免疫层析试纸条的研制

Study on Gold Immunochromatography Assay for Rapid Detection of Huperzine A

目的利用胶体金免疫层析技术建立一种快速检测石杉碱甲的方法。方法采用柠檬酸三钠还原法制备胶体金颗粒,标记抗石杉碱甲单克隆抗体并于4℃保存。石杉碱甲和羊抗鼠二抗分别结合于硝酸纤维膜上,依次将硝酸纤维素膜、金标垫、样品垫、吸水纸组装于PVC底板上,切割成胶体金试纸条。结果石杉碱甲快速检测试纸条的灵敏度为15μg·m L?1,检测时间为5 min,特异性高,批内和批间重复性良好。结论该法操作简单,使用方便,可用于现场快速检测石杉碱甲。

OBJECTIVE To establish a rapid assay for detection of huperzine A based on gold immunochromatography assay. METHODS Colloidal gold marked coupled with monoclonal antibody against huperzine A was jet sprayed onto the glass fiber and stored at 4 ℃. Huperzine A and anti-mouse goat immunoglobulins were jet-positioned onto a nitrocellulose membrane. Nitrocellulose membrane, gold combined pad, sample pad and absorbent paper were assembled and cut into detecting card, respectively. RESULTS The results showed that the sensitive of rapid for huperzine A visual detection limit was 15 μg·m L^-1 and detection time within 5 min. Repeatability and specificity of strip was good. CONCLUSION It is very simple and convenient to rapidly detect huperzine A on the spot.