期刊
MICROCHIMICA ACTA
卷 186, 期 8, 页码 -出版社
SPRINGER WIEN
DOI: 10.1007/s00604-019-3666-4
关键词
Hairpin probe; Rolling circle amplification; Hybridization chain reaction; Intracellular assay; DNAzyme
资金
- National Natural Science Foundation of China [21775122, 21775124, 51473136, 21575116, 21705115]
- Natural Science Foundation of Chongqing City China [cstc2018jcyjAX0693]
- Key Laboratory of Sensor Analysis of Tumor Marker, Ministry of Education, QingdaoUniversity of Science and Technology [SATM201604]
A DNA dendrimer was constructed by combination of rolling circle amplification (RCA) and hybridization chain reaction (HCR). A fluorescence resonance energy transfer (FRET) pair consisting of 6-carboxyfluorescein (FAM) and 5-carboxytetramethylrhodamine (TAMRA) was then created as a signaling probe for the ultrasensitive detection of Pb2+. Firstly, the DNAzyme released by Pb2+-induced recycling amplification as a primer induces RCA to open two different hairpins for generating repeated Y structures. Next, two other hairpins (labeled with FAM and TAMRA, respectively) are opened by the Y structures to trigger the HCR. As a result, a DNA dendrimer is generated. It is high loading with FRET pair and also promotes FRET pair mutually close to produce a remarkable FRET signal. Hence, ultrasensitive detection of Pb2+ is accomplished by measurement of the ratio of the yellow fluorescence of TAMRA (peakingat 588nm) and the green fluorescence of FAM (at 525nm). The method works in the 0.001 to 10nM Pb2+ concentration range and has a 0.3 pM detection limit. It was applied to determination of intracellular Pb2+ with convincing performance.
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