一种用于农业和水产养殖产品中 AFB 检测的高灵敏度和组特异性酶联免疫吸附测定（ELISA）。
A Highly Sensitive and Group-Specific Enzyme-Linked Immunosorbent Assay (ELISA) for the Detection of AFB in Agriculture and Aquiculture Products.
机构信息
The Key Lab of Health Chemistry and Molecular Diagnosis of Suzhou, College of Chemistry, Chemical Engineering and Materials Science, Soochow University, Renai Road 199, Suzhou 215123, China.
School of Biology & Basic Medical Science, Soochow University, Renai Road 199, Suzhou 215123, China.
出版信息
Molecules. 2024 May 12;29(10):2280. doi: 10.3390/molecules29102280.
Aflatoxins (AFs) including AFB, AFB, AFG and AFG are widely found in agriculture products, and AFB is considered one of the most toxic and harmful mycotoxins. Herein, a highly sensitive (at the pg mL level) and group-specific enzyme-linked immunosorbent assay (ELISA) for the detection of AFB in agricultural and aquiculture products was developed. The AFB derivative containing a carboxylic group was synthesized and covalently linked to bovine serum albumin (BSA). The AFB-BSA conjugate was used as an immunogen to immunize mice. A high-quality monoclonal antibody (mAb) against AFB was produced by hybridoma technology, and the mAb-based ELISA for AFB was established. IC and limit of detection (LOD) of the ELISA for AFB were 90 pg mL and 18 pg mL, respectively. The cross-reactivities (CRs) of the assay with AFB, AFG, and AFG were 23.6%, 42.5%, and 1.9%, respectively, revealing some degree of group specificity. Corn flour, wheat flour, and crab roe samples spiked with different contents of AFB were subjected to ELISA procedures. The recoveries and relative standard deviation (RSD) of the ELISA for AFB in spiked samples were 78.3-116.6% and 1.49-13.21% ( = 3), respectively. Wheat flour samples spiked with the mixed AF (AFB, AFB, AFG, AFG) standard solution were measured by ELISA and LC-MS/MS simultaneously. It was demonstrated that the proposed ELISA can be used as a screening method for evaluation of AFs (AFB, AFB, AFG, AFG) in wheat flour samples.
黄曲霉毒素（AFs）包括 AFB1、AFB2、AFG1 和 AFG2，广泛存在于农产品中，其中 AFB1 被认为是毒性和危害性最大的真菌毒素之一。本研究建立了一种用于检测农产品和水产品中 AFB1 的高灵敏度（pg/mL 级）和组特异性酶联免疫吸附测定法（ELISA）。合成了含有羧基的 AFB1 衍生物，并将其与牛血清白蛋白（BSA）共价连接。将 AFB1-BSA 缀合物用作免疫原免疫小鼠。通过杂交瘤技术生产了针对 AFB1 的高质量单克隆抗体（mAb），并建立了基于 mAb 的 AFB1 ELISA。该 ELISA 对 AFB1 的 IC50 和检测限（LOD）分别为 90 pg/mL 和 18 pg/mL。该检测方法对 AFB1、AFG1 和 AFG2 的交叉反应率（CR）分别为 23.6%、42.5%和 1.9%，显示出一定程度的组特异性。用不同含量 AFB1 对玉米粉、小麦粉和蟹黄样品进行 ELISA 检测。ELISA 检测 AFB1 加标样品的回收率和相对标准偏差（RSD）分别为 78.3%-116.6%和 1.49%-13.21%（n=3）。同时用 ELISA 和 LC-MS/MS 法对小麦粉样品中混合 AF（AFB1、AFB2、AFG1、AFG2）标准溶液进行了测定。结果表明，该 ELISA 可作为评价小麦粉样品中 AFs（AFB1、AFB2、AFG1、AFG2）的筛选方法。
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