SDR-ELISA: Ultrasensitive and high-throughput nucleic acid detection based on antibody-like DNA nanostructure.
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-digoxigenin 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 colorimetric 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.
Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, China; Guangdong Key Laboratory of Agricultural Environment Pollution Integrated Control, Guangdong Institute of Eco-Environmental and Soil Sciences, Guangzhou 510650, China; University of Chinese Academy of Sciences, Beijing, China.
Junlin
J
Wen
Guangdong Key Laboratory of Agricultural Environment Pollution Integrated Control, Guangdong Institute of Eco-Environmental and Soil Sciences, Guangzhou 510650, China.
Junhua
J
Chen
Guangdong Key Laboratory of Agricultural Environment Pollution Integrated Control, Guangdong Institute of Eco-Environmental and Soil Sciences, Guangzhou 510650, China.
Li
L
Zhuang
Guangdong Key Laboratory of Agricultural Environment Pollution Integrated Control, Guangdong Institute of Eco-Environmental and Soil Sciences, Guangzhou 510650, China. Electronic address: sgzhou@soil.gd.cn.
Shungui
S
Zhou
D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D015374:Biosensing Techniques; D001710:Biotin / Q000737:chemistry; D004247:DNA / Q000737:chemistry* / Q000276:immunology; D004076:Digoxigenin / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D006735:Horseradish Peroxidase / Q000737:chemistry; D009696:Nucleic Acids / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D019809:Streptavidin / Q000737:chemistry
D000906:Antibodies; D009696:Nucleic Acids; D001710:Biotin; D004247:DNA; D019809:Streptavidin; D006735:Horseradish Peroxidase; D004076:Digoxigenin
Colorimetric assay; Enzyme linked immunosorbent assay; High-throughput; Shewanella oneidensis; Strand displacement reaction