College of Environmental Science and Engineering/Sino-Canada R&D Centre on Water and Environmental Safety, Nankai University, Tianjin 300071, China E-mail:
Environmental Applied Science and Management, Ryerson University, 350 Victoria Street, Toronto, Ontario, M5B 2K3, Canada.
Water Sci Technol. 2022 Feb;85(3):900-913. doi: 10.2166/wst.2021.640.
The enzyme-linked immunosorbent assay (ELISA), as a universal method for the determination of Microcystins, is of great significance for the rapid detection of Microcystins pollution. This study aimed to propose a simplified validation method for Microcystins ELISA kit by summarizing related documents and guidelines. After summarizing and clarifying from 20 validation parameters, 11 parameters were selected to simplify the validation of Microcystins ELISA kit. In addition, the acceptable range and validation details of each parameter were analyzed. The results indicated that the coefficient of determination of the Microcystin-LR standard curve was higher than 0.99. The concentration of quality control samples was within control limits. The accuracy of spiked and proficient samples was within 70%-130%. The variability of intra-assay, inter-assay, and reproducibility was less than 11, 15 and 21%, respectively. The LOD and LLOQ were 0.002 μg/L and 0.05 μg/L, respectively. When the concentration of Microcystins exceeded 5 μg/L, it was recommended to dilute the samples to the working range before detection. The specificity was estimated with seven Microcystin analogues and three amino acids, indicating that the cross-reactivity was less than 30%. These results revealed that the ELISA kit was satisfactory for detecting Microcystins in water.
酶联免疫吸附测定法（ELISA）作为一种测定微囊藻毒素的通用方法，对于快速检测微囊藻毒素污染具有重要意义。本研究旨在通过总结相关文献和指南，提出一种简化微囊藻毒素 ELISA 试剂盒验证方法。在对 20 个验证参数进行总结和澄清后，选择了 11 个参数来简化微囊藻毒素 ELISA 试剂盒的验证。此外，还分析了每个参数的可接受范围和验证细节。结果表明，微囊藻毒素-LR 标准曲线的决定系数高于 0.99。质控样品的浓度在控制范围内。加标和熟练样品的准确度在 70%-130%之间。内、间和再现性的变异系数分别小于 11%、15%和 21%。LOD 和 LLOQ 分别为 0.002 μg/L 和 0.05 μg/L。当微囊藻毒素浓度超过 5 μg/L 时，建议在检测前将样品稀释至工作范围。用七种微囊藻毒素类似物和三种氨基酸评估特异性，表明交叉反应性小于 30%。这些结果表明，该 ELISA 试剂盒可用于检测水中的微囊藻毒素。
