集成免疫磁珠富集和免疫层析的黄曲霉毒素M_1快速检测法

Rapid Detection of Aflatoxin M_1 by Immunochromatography Combined with Enrichment Based on Immunomagnetic Nanobeads

以喷涂了检测抗原黄曲霉毒素M1-BSA和驴抗鼠二抗形成检测线和质控线的硝酸纤维膜制备免疫层析试纸条,采用EDC/NHS法制备偶联了抗黄曲霉毒素M1单克隆抗体的免疫磁珠。免疫磁珠与待检样本混合,经捕获、磁分离后,浓缩重悬液直接用免疫层析试纸条检测,首次建立了集浓缩样本与免疫层析于一体的黄曲霉毒素M1快速检测法。该方法用于检测原料乳中黄曲霉毒素M1,检出限为0.1μg/L,低于我国制定的黄曲霉毒素M1限量标准(0.5μg/L),与其它真菌毒素和原料乳中常检违法添加物无交叉反应,分析结果与酶联免疫吸附法(ELISA)结果一致。本方法适合现场快速检测原料乳中黄曲霉毒素M1。

Immunochromatographic strip was assembled with conjugate pad, sample pad, absorbent pad, and the nitrocellulose membrane on which aflatoxin M1-BSA and donkey anti-mouse antibody was sprayed and served as test line and control line, respectively. Anti-aflatoxin M1 coated immunomagnetic nanobeads was synthesized by an EDC/NHS method. Immunomagnetic nanobeads were mixed with sample for capturing and separation. The enrichment sample was detected by immunochromatographic strip. A rapid detection method of aflatoxin M1 containing sample enrichment and immunochromatographic assay was firstly set up. The limit of detection of this assay for aflatoxin M1 in raw milk was 0. 1 μg/L, which was lower than the legal limit of 0. 5 μg/L set by China. No cross-reactions were found with other mycotoxins and common illegal additives. The result of the method was in good agreement with that of ELISA. The assay can be applied for fast detection of aflatoxin M1 on-site in raw milk.