Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan, 430062, China; Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Wuhan, 430062, China; Laboratory of Quality & Safety Risk Assessment for Oilseed Products (Wuhan), Ministry of Agriculture, Wuhan, 430062, China; Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan, 430062, China; Quality Inspection & Test Center for Oilseed Products, Ministry of Agriculture, Wuhan, 430062, China.
Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan, 430062, China; Laboratory of Quality & Safety Risk Assessment for Oilseed Products (Wuhan), Ministry of Agriculture, Wuhan, 430062, China; Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan, 430062, China.
Anal Chim Acta. 2019 Oct 31;1078:142-150. doi: 10.1016/j.aca.2019.06.015. Epub 2019 Jun 10.
Mycotoxins and pesticides are prevalent in cereal food. It is difficult to detect these two kinds of hazard factors simultaneously in rapid assay. In order to find a solution to the problem, carbamates and aflatoxins were selected in this study to establish a rapid, on-site, and quantitative paper sensor. Two novel monoclonal antibodies (mAbs) against carbaryl and carbofuran (1D2 and G11) were developed. The IC50 values (half maximal inhibitory concentration) were 0.8 ng/mL and 217.6 ng/mL for carbaryl and carbofuran, respectively. Based on the sensitive and specific mAbs, a multi-TRFICA (time-resolved fluorescence) paper sensor was developed, which simultaneously detected six types of hazardous chemicals, including AFB, AFB, AFG, AFG, carbaryl, and carbofuran. A universal sample pretreatment method for mycotoxins and pesticides was explored to apply on established competitive indirect enzyme-linked immunosorbent assay (icELISA) and multi-TRFICA-paper sensor. The established paper sensor can be easily observed with naked eyes, qualitatively under a UV lamp, and quantitated using a home-made device. It exhibited a calculated limit of quantity for AFTs, carbaryl, and carbofuran of 0.03, 0.02, and 60.2 ng/mL in corn samples, respectively. The spiking-recoveries and real sample studies proved that multi-TRFICA-paper sensor is an accurate, sensitive, and high throughput detection method for simple and low-cost analysis in corn samples.
真菌毒素和农药普遍存在于谷物食品中。在快速检测中，同时检测这两种危害因素具有一定难度。为了解决这个问题，本研究选择了氨基甲酸酯类农药和黄曲霉毒素来建立一种快速、现场、定量的纸质传感器。本研究开发了两种针对甲萘威和克百威的新型单克隆抗体（mAb）（1D2 和 G11）。甲萘威和克百威的 IC50 值（半抑制浓度）分别为 0.8ng/mL 和 217.6ng/mL。基于这两种高灵敏度和高特异性的 mAb，开发了一种多时间分辨荧光（TRF）的纸质传感器，可同时检测六种危险化学品，包括 AFB1、AFB2、AFG1、AFG2、甲萘威和克百威。本研究还探索了一种适用于建立竞争间接酶联免疫吸附试验（icELISA）和多-TRFICA 纸质传感器的真菌毒素和农药通用样品预处理方法。建立的纸质传感器可通过肉眼观察、在紫外灯下定性观察和使用自制设备定量观察。该纸质传感器对玉米样品中 AFTs、甲萘威和克百威的计算定量限分别为 0.03、0.02 和 60.2ng/mL。加标回收率和实际样品研究证明，多-TRFICA 纸质传感器是一种准确、灵敏、高通量的检测方法，可用于玉米样品的简单、低成本分析。
