Department of Applied Chemistry, Osaka Prefecture University, 1-2 Gakuen, Naka, Sakai, Osaka, 599-8570, Japan.
GreenChem. Inc., 19-19 Tsuruta, Nishi, Sakai, Osaka 593-8323, Japan.
Anal Sci. 2020 Dec 10;36(12):1461-1465. doi: 10.2116/analsci.20P225. Epub 2020 Aug 7.
Enzyme-linked immunosorbent assay (ELISA) is a widespread analytical biochemistry assay. In this work, a direct ELISA method using a metallic nanoparticle (NP)-immobilized 96-well plate was developed for high-throughput, highly sensitive fluorescence analysis. Immobilization of metallic NPs on a 96-well plate effectively amplified fluorescence signals of the assay. The silver (Ag) NP-immobilized plate showed the best fluorescence enhancement effect of all the metal-immobilized plates tested. We used the Ag NP-immobilized plate to detect biomolecules and bacteria and found that both the fluorescence intensity and the limit of detection (LOD) were strongly enhanced by more than 100 times compared with those of the unmodified 96-well plates. Quantitative and qualitative considerations for target bacteria regarding the impact of autofluorescence on detection were successfully obtained for several strains. Our results demonstrate the potential of applying Ag NPs for enhancing the efficiency of direct and indirect ELISA assays.
酶联免疫吸附测定（ELISA）是一种广泛应用的分析生物化学检测方法。在这项工作中，我们开发了一种使用金属纳米粒子（NP）固定的 96 孔板的直接 ELISA 方法，用于高通量、高灵敏度的荧光分析。将金属 NPs 固定在 96 孔板上可以有效地放大检测的荧光信号。在所有测试的金属固定化板中，银（Ag）NP 固定化板显示出最佳的荧光增强效果。我们使用 Ag NP 固定化板来检测生物分子和细菌，发现与未修饰的 96 孔板相比，荧光强度和检测限（LOD）都增强了 100 多倍。对于几种菌株，我们成功地从定量和定性方面考虑了目标细菌的自荧光对检测的影响。我们的结果表明，Ag NPs 在提高直接和间接 ELISA 检测效率方面具有应用潜力。
