人参皂苷Re胶体金免疫检测试纸条的制备与应用

Development of a lateral flow dipstick immunoassay for rapid detection of ginsenoside Re

利用胶体金标记的经饱和硫酸铵纯化的人参皂苷Re单克隆抗体、人参皂苷Re-牛血清白蛋白(GRe-BSA)、羊抗鼠IgG制备了人参皂苷Re胶体金免疫检测试纸条。该试纸条的最低检测极限为200μg·L-1。检测时间10 min。取10 mg不同年份的人参样品分别用5 mL自来水提取30 min,分别用试纸条和酶联免疫检测法(indirect competitive enzyme-linked im-munosorbent assay,icELISA)检测。结果为:直接用试纸条测定上清液可区别真伪人参;上清液稀释100倍,用试纸条可区分出人参质量优劣。试纸条检测结果和酶联免疫检测方法测定结果完全一致。所制备的胶体金免疫检测试纸条可以用于人参的快速真伪鉴别和质量优劣判断。

A sensitive antibody-based lateral flow dipstick was developed for ginsenoside Re （GRe） detection. The stick con- sisted of a sample pad, a conjugate pad, membrane and an absorbent pad. The membrane was coated with two capture reagents, GRe- BSA conjugate and goat anti-mouse antibodies, forming a test line and a control line, respectively. The conjugate pad was saturated with colloidal gold particles coated with affinity purified monoclonal anti-GRe antibody. The visual detection limit was 200μg·L^-1 of GRe and the reaction time was 10 min. The Panax ginseng roots were identified after these samples （ 10 mg） were extracted with 5 mL tap water for 30 min at room temperature, and the extracts were tested by the dipsticks and ELISA kit. The true and false P. ginseng could be distinguished with dipsticks. The dipstick could be used to detect the quality of the P. ginseng samples when the extract was diluted 100-folds. The results were compared with those obtained using an indirect competitive enzyme-linked immunosorbent assay （icELISA）. The dipstick assay proved to be a sensitive and rapid tool for quality control of P. ginseng.