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基于硫掺杂石墨相氮化碳量子点与 AgS 纳米晶偶联的流感病毒荧光免疫分析。

Fluoroimmunoassay of influenza virus using sulfur-doped graphitic carbon nitride quantum dots coupled with AgS nanocrystals.

机构信息

Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka, 422-8529, Japan.

Research Institute of Electronics, Shizuoka University, 3-5-1 Johoku, Nakaku, Hamamatsu, Shizuoka, 432-8011, Japan.

出版信息

Mikrochim Acta. 2020 Jul 21;187(8):466. doi: 10.1007/s00604-020-04433-1.

Abstract

Novel sulfur-doped graphitic carbon nitride quantum dots (S-gCNQDs) are synthesized using a single-source precursor in a one-step solvothermal process. The S-gCNQDs with a size of ~ 5-nm displayed a strong green intrinsic fluorescence at 512 nm when excited at 400 nm, with a quantum yield of ~ 33% in aqueous solution. The prepared S-gCNQDs and AgS nanocrystals were applied as innovative functional materials to fabricate a biosensor for virus detection based on the conjugation of specific anti-human influenza A monoclonal antibody to the S-gCNQDs and AgS NCs, respectively. In the presence of the influenza A virus, an interaction between the S-gCNQDs/AgS-labeled antibody resulted in the formation of a nanosandwich structure, which is accompanied by the fluorescence enhancement of the S-gCNQDs. The change in fluorescence intensity linearly correlats with the concentration of the influenza A virus (H1N1) in the 10 fg/mL to 1.0 ng/mL range, with a limit of detection of 5.5 fg/mL. The assay was applied to the assay of clinically isolated influenza A virus (H3N2/Yokohama) mixed with human serum. The obtained limit of detection was 100 PFU/mL within the detection range of 10- 5 × 10 PFU/mL for the H3N2 virus. Graphical abstract.

摘要

新型硫掺杂石墨相氮化碳量子点(S-gCNQDs)是通过一步溶剂热法使用单源前体制备的。在 400nm 激发下,尺寸约为 5nm 的 S-gCNQDs 在 512nm 处显示出强烈的绿色本征荧光,在水溶液中的量子产率约为 33%。所制备的 S-gCNQDs 和 AgS 纳米晶分别作为创新的功能材料,用于基于特异性抗人流感 A 单克隆抗体与 S-gCNQDs 和 AgS NCs 的偶联来构建病毒检测的生物传感器。在流感 A 病毒存在下,S-gCNQDs/AgS 标记抗体之间的相互作用导致形成纳米三明治结构,同时 S-gCNQDs 的荧光增强。荧光强度的变化与流感 A 病毒(H1N1)在 10 fg/mL 至 1.0 ng/mL 范围内的浓度呈线性相关,检测限为 5.5 fg/mL。该测定法应用于临床分离的流感 A 病毒(H3N2/Yokohama)与人血清的混合测定。对于 H3N2 病毒,在 10-5×10 PFU/mL 的检测范围内,获得的检测限为 100 PFU/mL。

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