An enzyme-linked immunosorbent assay and a gold-nanoparticle based immunochromatographic test for amatoxins using recombinant antibody

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• 作者：Kuo He ; Xiuyuan Zhang ; Ruiping Zhao ; Lixia Wang ; Tingting Feng…
• 刊名：Microchimica Acta
• 出版年：2016
• 出版时间：July 2016
• 年：2016
• 卷：183
• 期：7
• 页码：2211-2219
• 全文大小：1,003 KB
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Analytical Chemistry
  Inorganic Chemistry
  Physical Chemistry
  Characterization and Evaluation Materials
  Monitoring, Environmental Analysis and Environmental Ecotoxicology
  
• 出版者：Springer Wien
• ISSN：1436-5073
• 卷排序：183

文摘

The authors describe two kinds of rapid assays for the determination of amatoxins in mushrooms. The first is an enzyme-linked immunosorbent assay (ELISA) using horseradish peroxidase. The second is a rapid immunochromatographic assay that uses colloidal gold as a red label (CG-ICA). Both are based on the use of a well-characterized recombinant single chain variable fragment antibody (named scFv-A4). The half-maximum inhibition concentrations (IC50) of α-amanitin, β-amanitin and γ-amanitin are 78, 85 and 90 ng⋅mL‾^{1, and the limits of detection (LODs; for IC15) are 1.9, 2.1 and 2.8 ng⋅mL‾1. The method was applied to the determination of amanitins in mushrooms, and the LODs for α-amanitin, β-amanitin and γ-amanitin in mushroom samples were found to be 4.9, 6.4 and 8.3 ng⋅mL‾1. The visual minimum detection limits of the optimized CGIA are 4 and 6 ng⋅mL‾1 for mushroom samples. The test can be performed within 10 min. The results of the analysis of spiked samples showed that the CG-IA can rapidly and semi-quantitatively quantify amatoxins in mushroom samples on site and at low costs.}