International Joint Research Laboratory for Biointerface and Biodetection, School, of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, 214122, China.
Key Laboratory of Food Safety Rapid Detection Technology and Product Evaluation for Market Regulation of Jiangsu Province, Suzhou, Jiangsu, 215133, China.
Anal Methods. 2023 Jul 13;15(27):3353-3361. doi: 10.1039/d3ay00562c.
Due to its unique insecticidal and acaricidal mechanism of action, and ability to mix with most insecticides and fungicides, diafenthiuron (DIAF) is widely used in the cultivation of fruits and vegetables. However, this insecticide can cause unacceptable harm to organisms, making the detection of DIAF residues in fruits and vegetables crucial. In this study, a novel hapten based on the structure of DIAF was utilized to prepare a monoclonal antibody (mAb) with high specificity and sensitivity. The half maximum inhibitory concentration (IC) of the anti-DIAF mAb was 20.96 μg kg as determined by ic-ELISA and little cross-reactivity with other analogues. Next, a GNP-based lateral flow immunoassay (LFIA) was developed to detect DIAF in cabbages and apples. The optimized LFIA, for cabbage samples, showed a visual limit of detection (vLOD), cut-off value and calculated limit of detection (cLOD) of 0.1 mg kg, 10 mg kg and 1.5 μg kg, respectively, and for apples 0.1 mg kg, 5 mg kg and 3.4 μg kg, respectively. Recovery rates in cabbage and apples were 89.4-105.0% and 105.3-112.0%, with a coefficient of variation of 2.73-5.71% and 2.15-7.56%, respectively. These results indicated that the established LFIA based on our anti-DIAF mAb was a reliable method for rapid detection of DIAF in cabbage and apple samples.
由于其独特的杀虫和杀螨作用机制,以及能够与大多数杀虫剂和杀菌剂混合使用,噻苯隆(DIAF)在水果和蔬菜的种植中被广泛应用。然而,这种杀虫剂会对生物体造成不可接受的危害,因此检测水果和蔬菜中的 DIAF 残留至关重要。在本研究中,利用基于 DIAF 结构的新型半抗原制备了具有高特异性和灵敏度的单克隆抗体(mAb)。ic-ELISA 测定的抗 DIAF mAb 的半最大抑制浓度(IC)为 20.96 μg kg,与其他类似物的交叉反应性很小。接下来,基于 GNP 的侧向流动免疫分析(LFIA)被开发用于检测白菜和苹果中的 DIAF。优化后的 LFIA 用于检测白菜样品,其可视检测限(vLOD)、截止值和计算检测限(cLOD)分别为 0.1 mg kg、10 mg kg 和 1.5 μg kg,用于检测苹果样品,其 vLOD、截止值和 cLOD 分别为 0.1 mg kg、5 mg kg 和 3.4 μg kg。白菜和苹果中的回收率分别为 89.4-105.0%和 105.3-112.0%,变异系数分别为 2.73-5.71%和 2.15-7.56%。这些结果表明,基于我们的抗 DIAF mAb 建立的 LFIA 是一种快速检测白菜和苹果样品中 DIAF 的可靠方法。