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

文献类型： 外文期刊

作者： Wen, Junlin 1 ; Chen, Junhua 2 ; Zhuang, Li 2 ; Zhou, Shungui 2 ;

作者机构： 1.Chinese Acad Sci, Guangzhou Inst Geochem, Beijing 100864, Peoples R China

2.Guangdong Inst Ecoenvironm & Soil Sci, Guangdong Key Lab Agr Environm Pollut Integrated, Guangzhou 510650, Guangdong, Peoples R China

3.Univ Chinese Acad Sci, Beijing, Peoples R China

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 ）

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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.

  • 相关文献

[1]Axial Ligation of Heme in c-Type Cytochromes of Living Shewanella oneidensis: A New Insight into Enhanced Extracellular Electron Transfer. Yuan, Yong,Zhou, Shungui,Li, Laicai. 2015

[2]Designed diblock hairpin probes for the nonenzymatic and label-free detection of nucleic acid. Wen, Junlin,Wen, Junlin,Chen, Junhua,Zhuang, Li,Zhou, Shungui,Wen, Junlin.

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