Colorimetric biosensor using dual-amplification of enzyme-free reaction through universal hybridization chain reaction system.

Authors:
Won Jong Rhee
Won Jong Rhee
Seoul National University
South Korea
Kyu Won Kim
Kyu Won Kim
Seoul National University College of Medicine
Byeong Hee Hwang
Byeong Hee Hwang
Pohang University of Science and Technology

Biotechnol Bioeng 2019 Jul 17;116(7):1567-1574. Epub 2019 Apr 17.

Department of Bioengineering and Nano-bioengineering, Incheon National University, Incheon, Korea.

On-site genetic detection needs to develop a sensitive and straightforward biosensor without special equipment, which can detect various genetic biomarkers. Hybridization chain reaction (HCR) amplifying signal isothermally could be considered as a good candidate for on-site detection. Here, we developed a novel genetic biosensor on the basis of enzyme-free dual-amplification of universal hybridization chain reaction (uHCR) and hemin/G-quadruplex horseradish peroxidase (HRP)-mimicking DNAzyme. The uHCR is the strategy which enables simple design for multiple target detection by the introduction of target-specific trigger hairpin without changing the whole system according to a target change. Also, HRP-mimicking DNAzyme could produce a sensitive and quantitative colorimetric signal with increased stability with a limit of detection (LOD) of 5.67 nM. The universality of the uHCR biosensor was proven by the detection of four different targets (miR-21, miR-125b, KRAS-Q61K, and BRAF-V600E) for cancer diagnosis. The uHCR biosensor showed specificity that could discriminate single-nucleotide polymorphism. Moreover, the uHCR biosensor could detect targets in the diluted serum sample. Overall, the uHCR biosensor demonstrated the potential for field testing with a simple redesign without complicated steps or special equipment using a universal hairpin system and enzyme-free amplification. This strategy could enable stable and sensitive detection of a variety of targets. Therefore, it could be applied to urgent detection of various pathogens, remote diagnosis, and self-screening of diseases.

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Source
http://dx.doi.org/10.1002/bit.26978DOI Listing
July 2019
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