毒死蜱等10种农药多残留快速检测芯片研究

Immunochip Assay for Multi-residue Rapid Detection of Ten Kinds of Common Pesticides

以醛基修饰的载玻片为固相载体,农药竞争抗原为包被抗原,胶体金为标记材料,抗农药单克隆抗体为识别元件,银增强试剂用于放大信号,建立了一种含有10对包被抗原与农药抗体组合的免疫芯片,可同时检测农产品中毒死蜱、三唑磷、克百威、噻虫啉、吡虫啉、多菌灵、异菌脲、涕灭威、甲氰菊酯和百菌清共10种农药。对胶体金标记探针、包被抗原与农药抗体组合等免疫芯片反应条件进行了优化。本方法对10种农药的检出限(IC 20 )达到1.49~15.72 μg/L,检测仅需1.5 h。将此芯片方法应用于水果(苹果)、蔬菜(黄瓜)以及茶叶(红茶)中的10种农药残留检测,其最低检出量可满足相应作物中的农药最大残留限量(MRL)检测要求,且此芯片检测方法具有较高的准确度和精密度,农药的添加回收率为82.1%~120.8%,批内相对标准偏差RSD≤10.4%,批间RSD≤12.1%,与质谱仪检测结果的相关系数大于0.9591。本研究为农药多残留快速检测分析提供了技术支持,具有良好的实用价值。

With aldehyde-modified glass slide as solid phase, pesticide competitive antigens as coating antigens, gold nanoparticles as labeling material, anti-pesticide monoclonal antibodies as recognition elements and silver-enhance reagent as signal amplification, an immunochip containing 10 combinations of coating antigens and antibodies against corresponding pesticides was developed, which could be used for simultaneous detection of 10 kinds of pesticide residues in agricultural products, including chlorpyrifos, triazophos, carbofuran, thiazoline, imidacloprid, carbendazim, iprodione, aldicarb, fenpropathrin and chlorothalonil. The limits of detection (IC 20 ) of 10 kinds of pesticides were 1.49-15.72 μg/L under the optimized reaction conditions of immunochip. Moreover, the detection process took only 1.5 h. The chip method was applied to the detection of 10 kinds of pesticide residues in fruits (apples), vegetables (cucumbers) and tea (black tea). The sensitivity of the chip met the national Maximum Residue Limit (MRL) standard of pesticides on corresponding crops. Furthermore, the immunochip assay had high accuracy and precision, with recoveries of pesticides from 82.1% to120.8%, and intra-day and inter-day coefficient of variation less than 10.4% and 12.1%, respectively. The results from the immunochip assay had good correlation with those from mass spectrometer, with R 2 of over 0.9591. This immunochip assay provided technical support for rapid detection of pesticide residues, and had practical significance for guaranteeing the quality and safety of agricultural products.