The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, Fujian Key Laboratory of Pathogenic Fungi and Mycotoxins, and School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
The Ministry of Education Key Laboratory of Biopesticide and Chemical Biology, Fujian Key Laboratory of Pathogenic Fungi and Mycotoxins, and School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
Food Chem. 2020 Jul 30;319:126505. doi: 10.1016/j.foodchem.2020.126505. Epub 2020 Feb 27.
The high content of Penicillic acid (PA) in the feed pose threat to human health and cause serious losses to economic wealth through the enrichment effect of the food chain. The reliable and rapidly detection of PA is of significant importance to ensure food safety. In this study, indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and immunochromatographic test strips (ICTS) were established for PA determination based on anti-PA mAb secreted by 4H9 cell line. The linear range of ic-ELISA detection was 0.12-1.95 μg/mL, and the limit of detection (LOD) was 0.03 μg/mL. Then, conventional gold nanospheres (AuNS) with the average diameter of 20 nm were synthetized and AuNS-based strip was developed for rapidly detection of PA. The visual LOD (vLOD) of the AuNS-based strip was 3.9 μg/mL and the assay time of visual evaluation was less than 10 min without any instrument. To enhance the signal sensitivity of the ICTS, the larger size (about 85 nm) of gold nanoflowers (AuNFs) was prepared in our work, and was used as higher signal reporter to establish the AuNF-based strip for PA determination. Fortunately, the vLOD of AuNF-based strip was 0.97 μg/mL, which was approximately 4-fold lower than that of traditional AuNS-based strip. In summary, the rapid and sensitive immunoassays established in this study could be applied to detect and analyze the contamination of PA toxin in real food samples.
饲料中较高含量的青霉素酸(PA)通过食物链的富集效应对人类健康构成威胁,并造成严重的经济财富损失。可靠且快速地检测 PA 对于确保食品安全具有重要意义。在本研究中,基于 4H9 细胞系分泌的抗 PA mAb,建立了间接竞争酶联免疫吸附测定(ic-ELISA)和免疫层析测试条(ICTS)用于 PA 测定。ic-ELISA 检测的线性范围为 0.12-1.95μg/mL,检测限(LOD)为 0.03μg/mL。然后,合成了常规的平均直径为 20nm 的金纳米球(AuNS),并开发了基于 AuNS 的条带用于快速检测 PA。基于 AuNS 的条带的目视检测限(vLOD)为 3.9μg/mL,目视评估的测定时间不到 10 分钟,无需任何仪器。为了提高 ICTS 的信号灵敏度,我们在工作中制备了更大尺寸(约 85nm)的金纳米花(AuNFs),并将其用作更高信号报告器,以建立用于 PA 测定的基于 AuNF 的条带。幸运的是,基于 AuNF 的条带的 vLOD 为 0.97μg/mL,约比传统基于 AuNS 的条带低 4 倍。总之,本研究建立的快速灵敏免疫分析方法可用于检测和分析实际食品样品中 PA 毒素的污染。