Rahi Shraddha, Lanjekar Vikram, Ghormade Vandana
Nanobioscience Group, Agharkar Research Institute, GG Agarkar Road, Pune 411004, India.
Bioenergy Group, Agharkar Research Institute, GG Agarkar Road, Pune 411004, India.
Food Chem. 2023 Jul 1;413:135651. doi: 10.1016/j.foodchem.2023.135651. Epub 2023 Feb 10.
Aflatoxin B1 (AFB1) is a hepatotoxic and carcinogenic food contaminant. Although on-site paper-based detection is sensitive it depends on expensive antibodies which are difficult to raise against mycotoxins. Here, we rationally designed a high binding octapeptide, N-KSGKSKPR-C peptide for AFB1 detection, by molecular docking, as confirmed by indirect ELISA (K 323 nM). Further, conjugation of octapeptide with gold nanoparticles (26 nm) permitted its use as a visual detection agent in rapid, sensitive dot-blot assay (LOD 0.39 μg/kg). The assay displayed negligible cross-reactivity with co-contaminating mycotoxins. AFB1 recovery from spiked wheat sample was comparable by dot-blot (78-91 %) and HPLC (65-87 %). Evaluation of dot-blot using certified reference material and 146 food and feed samples showed high correlation R = 0.87 with HPLC. The assay displayed high accuracy (91 %), sensitivity (71 %) and specificity (96.5 %). Therefore, the developed dot-blot assay holds promise for monitoring AFB1 contamination in food and feed.
黄曲霉毒素B1（AFB1）是一种具有肝毒性和致癌性的食品污染物。尽管基于纸张的现场检测很灵敏，但它依赖于昂贵的抗体，而针对霉菌毒素产生这些抗体很困难。在此，我们通过分子对接合理设计了一种用于检测AFB1的高结合八肽N-KSGKSKPR-C肽，间接酶联免疫吸附测定（K 323 nM）证实了这一点。此外，八肽与金纳米颗粒（26 nm）的缀合使其能够用作快速、灵敏的斑点印迹测定中的可视化检测剂（检测限0.39 μg/kg）。该测定与共污染的霉菌毒素显示出可忽略不计的交叉反应性。通过斑点印迹法（78-91%）和高效液相色谱法（65-87%）从加标小麦样品中回收AFB1的结果相当。使用有证标准物质以及146份食品和饲料样品对斑点印迹法进行评估，结果显示与高效液相色谱法具有高度相关性，R = 0.87。该测定显示出高准确度（91%）、灵敏度（71%）和特异性（96.5%）。因此，所开发的斑点印迹测定法有望用于监测食品和饲料中的AFB1污染。
