Wu Shih-Wei, Ko Jiunn-Liang, Liu Biing-Hui, Yu Feng-Yih
Graduate Institute of Medicine, Chung Shan Medical University Taichung 40201 Taiwan.
Graduate Institute of Toxicology, College of Medicine, National Taiwan University Taipei 10051 Taiwan
RSC Adv. 2022 Jul 8;12(31):19981-19989. doi: 10.1039/d2ra02823a. eCollection 2022 Jul 6.
Citrinin (CTN) is a mycotoxin primarily produced by species. Excess consumption of CTN may lead to nephrotoxicity and hepatotoxicity. A pilot study for commercial production of competitive direct enzyme-linked immunosorbent assay (cdELISA) kit and an immunochromatographic strip (immunostrip) for screening CTN in red yeast rice is established in this study. The coating antibody and the CTN-horse radish peroxidase (HRP) concentrations were optimized to increase the sensitivity and specificity of cdELISA kit. The conjugation methods/ratios of CTN to HRP as well as the long-term stability of kit components were also evaluated. The IC and detection limit of the ELISA kit were determined to be 4.1 and 0.2 ng mL, respectively. Analysis of 20 red yeast rice samples using ELISA kits revealed the contamination levels of CTN from 64 to 29 404 ng g. The on-site rapid detection of CTN with the immunostrip showed that CTN levels in seven samples exceeded the regulatory limit of 5 ppm. Additionally, the coefficient correlation between the results of HPLC and ELISA kits of 20 samples was 0.96. Sensitive and convenient tools at commercial levels for detection of CTN contamination in food are established herein to protect the health of the public.
桔霉素（CTN）是一种主要由某些物种产生的霉菌毒素。过量摄入CTN可能导致肾毒性和肝毒性。本研究建立了一项用于商业生产竞争性直接酶联免疫吸附测定（cdELISA）试剂盒和免疫层析试纸条（免疫试纸）的初步研究，用于筛选红曲米中的CTN。优化了包被抗体和CTN-辣根过氧化物酶（HRP）的浓度，以提高cdELISA试剂盒的灵敏度和特异性。还评估了CTN与HRP的偶联方法/比例以及试剂盒组分的长期稳定性。ELISA试剂盒的半数抑制浓度（IC）和检测限分别确定为4.1和0.2 ng/mL。使用ELISA试剂盒对20个红曲米样品进行分析，结果显示CTN的污染水平为64至29404 ng/g。使用免疫试纸对CTN进行现场快速检测表明，7个样品中的CTN水平超过了5 ppm的监管限值。此外，20个样品的高效液相色谱（HPLC）结果与ELISA试剂盒结果之间的系数相关性为0.96。本文建立了商业水平上用于检测食品中CTN污染的灵敏且便捷的工具，以保护公众健康。
