期刊
MICROCHIMICA ACTA
卷 184, 期 10, 页码 4183-4188出版社
SPRINGER WIEN
DOI: 10.1007/s00604-017-2337-6
关键词
Isothermal nucleic acid amplification; Aptasensor; Fluorescence; Noble-metal nanoclusters; DNA-templated; Sensitive
资金
- National Key Basic Research Program [2011CB911000]
- National Natural Science Foundation of China [21527810, 21575036, 21190041, 21521063, 21505041]
- Graduate Student Research Innovation Project of Hunan Province [CX2016B118]
A fluorescence aptasensor is described that combines target-cycling strand displacement amplification (TCSDA) and synthesis of copper nanoclusters (CuNCs) templated with double-stranded DNA (dsDNA). Specifically, the detection scheme was applied to the determination of adenosine-5'-triphosphate (ATP) via target-induced structure switching design. The binding of an aptamer hairpin probe (AP) to ATP induces a structural switch from a hairpin shape to an open conformation. This facilitates hybridization with a primer and triggers a TCSDA reaction. This amplification step produces a large quantity of dsDNA that can directly act as a template for the synthesis of fluorescent CuNCs, thereby producing a strong red fluorescence (with excitation/emission maxima at 340/598 nm) that can be used to quantify ATP. The assay has a dynamic range that extends over 4 decades (from 0.01 nM to 100 nM) and a 5 pM detection limit. Conceivably, this detection scheme is applicable to numerous other analytes for which suitable aptamers are available.
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