SDR-ELISA: Ultrasensitive and high-throughput nucleic acid detection based on antibody-like DNA nanostructure

文献类型: 外文期刊

第一作者: Wen, Junlin

作者: Wen, Junlin;Wen, Junlin;Chen, Junhua;Zhuang, Li;Zhou, Shungui;Wen, Junlin

作者机构:

关键词: Calorimetric assay;High-throughput;Strand displacement reaction;Enzyme linked immunosorbent assay;Shewanella oneidensis

期刊名称:BIOSENSORS & BIOELECTRONICS ( 影响因子:10.618; 五年影响因子:9.323 )

ISSN:

年卷期:

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收录情况: SCI

摘要: An ultrasensitive and high-throughput nucleic acid detection system, termed as strand displacement reaction enzyme linked immunosorbent assay (SDR-ELISA), has been developed on the basis of antibody-like DNA nanostructures. Three digoxigenin or biotin modified hairpin probes are utilized to construct antibody-like DNA nanostructures that feature affinity toward streptavidin and anti-digmdgenin antibody via isothermal target triggered SDR amplification. These antibody-like nanostructures have been employed to conjugate horseradish-peroxidase-labeled anti-digoxigenin antibody with streptavidin that is immobilized on microliter plate wells for enzyme-linked calorimetric assay. The resulting SDR-ELISA system is ultrasensitive for target DNA with a low detection limit of 5 fM. Moreover, the SDR-ELISA system is capable of discriminating DNA sequences with single base mutations, and do so in a high-throughput manner by detection and quantification of up to 96 or 384 DNA samples in a single shot. This detection system is further applied to detect other DNA targets such as Shewanella oneidensis specific DNA sequence, which indicates the generality of proposed SDR-ELISA system. The integration of SDR amplification and convenient ELISA technique advances an intelligent strategy for ultrasensitive and high-throughput nucleic acid detection, which may be amenable for direct visual detection and quantification using an accompanying quantitative color chart.

分类号: Q

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