Simplified validation of the ELISA kit determination of Microcystins in surface water. (29th December 2021)

• Record Type: Journal Article
• Authors: Fan, Liang; Jeanne Huang, Jinhui; Lo, Ching Y.; Zhou, Bin; Fu, Xujin
• Is Part Of: Water science and technology. Volume 85:Number 3(2022)
• Page Start: 900
• Page End: 913
• Publication Date: 2021-12-29
• Subjects: acceptable range -- ELISA kit -- Microcystins -- simplified validation -- validation parameters; Water -- Pollution; Sewage -- Purification; Water quality management; Periodicals; 628.168
• DOI: 10.2166/wst.2021.640
• Languages: English
• ISSNs: 0273-1223

Abstract

Abstract: 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. HIGHLIGHTS: 20 validation parameters were summarized from related regulatory documents and studies. 11 parameters were selected for Microcystins ELISA kit validation after the clarification and simplification of summarized parameters. Acceptance range and validation details of selected parameters were investigated. A simplified method for Microcystins ELISA kit validation was proposed. Graphical Abstract