Key Laboratory of Food Quality and Safety of Jiangsu Province, Nanjing, 210014, China; College of Plant Protection, Nanjing Agricultural University, Nanjing, 210095, China.
Key Laboratory of Food Quality and Safety of Jiangsu Province, Nanjing, 210014, China.
Anal Biochem. 2023 Sep 15;677:115270. doi: 10.1016/j.ab.2023.115270. Epub 2023 Jul 31.
Bacillus thuringiensis (Bt) Cry toxins have been widely used in the development of genetically modified organisms (GMOs) for pest control. This work aimed to establish more cost effective methods for used Cry2Aa toxins. Three immunoassay methods (IC-ELISA, DAS-ELISA, and CLEIA) were successfully developed in this work. The mAb was used as the detecting antibody, for the IC-ELISA, the range of IC to IC was 1.11 μg/mL - 60.70 μg/mL, and an IC of 10.65 μg/mL. For the DAS-ELISA, the limit of detection (LOD) and limit of quantitation (LOQ) were 10.76 ng/mL and 20.70 ng/mL, respectively. For the CLEIA, the LOD and LOQ were 6.17 ng/mL and 7.40 ng/mL, respectively. The scFv-based detections were the most sensitive for detecting Cry2Aa. The LOD and LOQ for the DAS-ELISA were 118.75 ng/mL and 633.48 ng/mL, respectively. The LOD and LOQ for the CLEIA, read as 37.47 ng/mL and 70.23 ng/mL, respectively. The fact that Cry2Aa toxin was recovered in spiked grain samples further demonstrated that the approaches might be used to identify field samples. These methods provided good sensitivity, stability, and applicability for detecting Cry2Aa toxin, promising ultrasensitive monitoring and references for Cry toxins risk assessment.
苏云金芽孢杆菌（Bt）Cry 毒素已被广泛应用于害虫防治的转基因生物（GMO）的开发。本工作旨在建立更具成本效益的Cry2Aa 毒素使用方法。本工作成功建立了三种免疫分析方法（IC-ELISA、DAS-ELISA 和 CLEIA）。该单抗被用作检测抗体，对于 IC-ELISA，IC 范围为 1.11μg/mL-60.70μg/mL，IC 为 10.65μg/mL。对于 DAS-ELISA，检测限（LOD）和定量限（LOQ）分别为 10.76ng/mL 和 20.70ng/mL。对于 CLEIA，LOD 和 LOQ 分别为 6.17ng/mL 和 7.40ng/mL。基于 scFv 的检测方法对检测 Cry2Aa 最为敏感。DAS-ELISA 的 LOD 和 LOQ 分别为 118.75ng/mL 和 633.48ng/mL。CLEIA 的 LOD 和 LOQ 分别为 37.47ng/mL 和 70.23ng/mL。在添加的谷粒样品中回收 Cry2Aa 毒素的事实进一步证明，这些方法可用于鉴定田间样品。这些方法为检测 Cry2Aa 毒素提供了良好的灵敏度、稳定性和适用性，有望对 Cry 毒素风险评估进行超灵敏监测和参考。
