Journal
MICROCHIMICA ACTA
Volume 184, Issue 6, Pages 1603-1610Publisher
SPRINGER WIEN
DOI: 10.1007/s00604-017-2158-7
Keywords
Three way junction; Strand displacement; Colorimetric detection; Isothermal amplification
Categories
Funding
- National Natural Science Foundation of China [81271917]
- Natural Science Foundation of Zhejiang province [LY14H200002, LY15H200002, LY16H160023]
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The authors describe a method for DNA target recognition and signal amplification that is based on the target-induced formation of a three way junction. The subsequent assembly of two DNA probes releases the inhibitory strand and triggers a downstream strand displacement amplification. This causes the formation of a G-rich single sequence that binds to a hemin monomer with its peroxidase-mimicking properties. The resulting peroxidase (POx) activity is quantified by using H2O2 and TMB as the substrate. In the presence of an inhibitor, in contrast, the POx-like activity is strongly reduced. This forms the basis for a highly sensitive DNA assay. It has a 0.8 pM detection limit when operated at a wavelength of 450 nm and was applied to the isothermal determination of target DNA with high selectivity.
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