高灵敏的磁酶免疫吸附分析法用于谷物中伏马菌素FB_1的快速检测

RAPID DETECTION OF FUMONISIN B_1 BY THE SENSITIVE MAGNETIC ENZYME-LINKED IMMUNOASSAY

建立谷物中伏马菌素FB1磁酶免疫吸附分析法（Magnetic enzyme-linked immunoassay,MELISA）,并对实际样品进行测定。以自制磁性纳米颗粒为固相载体,负载伏马菌素FB1-卵清蛋白偶联物（FB1-OVA）,建立检测FB1的MELISA方法,并与传统ELISA法和国标方法（HPLC,GB/T 255228—2010）进行比较。该方法线性回归方程为y=-0.229 5x＋0.569 3（x为lg（CFB 1）,y为OD/OD0）,线性范围0.1~25.0 ng/m L,相关系数r=0.995 4,检测限为0.071 ng/m L,与其他4种真菌毒素交叉反应率均低于2.0%。在不同的加标水平下,玉米和小麦批内加标回收率分别为95.2%~120.0%和84.4%~120.0%,对应的批间加标回收率分别为90.0%~100.0%和90.0%~110.0%（其相应的批内和批间RSD分别小于6.5%和12.8%）。与传统的ELISA和HPLC法相比,MELISA具有更低的检测限和更高的灵敏度。本研究所建立的MELISA灵敏度高、操作简单、检测快速,适用于大批量谷物样品中FB1含量的快速测定。

The present study was to investigate the method of rapid detection of Fumonisin B1（FB1）in cereals by the sensitive magnetic enzyme-linked immunoassay（MELISA） and compared with the conventional ELISA and the National standard method（HPLC,GB/T 255228—2010）. The method was established with self-made magnetic nanoparticles as solid carrier and loading FB1-OVA artificial antigen. The linear range of the MELISA method for FB1 detection was from 0.1 ng/m L to 25.0 ng/m L. The equation of linear regression was y=-0.229 5x＋0.569 3（x,y were lg（CFB1）,and OD/OD0,respectively）,while the correlation coefficient was 0.995 4 and the detection limit was 0.071 ng/m L,and the cross-reactivity rates with other four mycotoxins were less than 2.0%.This method could be satisfactorily applied to determine FB1 in corn and wheat at three spiked levels（0.1,1.0,and 25 ng/m L） and the recoveries of intra-assays ranged from 95.2% to 120.0% for corn and 84.4% to 120.0%for wheat. For inter-assays,the recoveries ranged from 90.0% to 100.0% and 90.0% to 110.0% for corn and wheat,respectively. The relative standard deviations（RSD）of the recoveries were less than 6.5% and 12.8% for intra- and inter-assays. Compared with ELISA and HPLC,the MELISA had lower detection limit and higher sensitivity. The MELISA established in this study could be effectively used for the FB1 detection in cereals with the advantages of high sensitivity and specificity,simple operation,rapid detection and high-throughput analysis.