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Colorimetric Biosensor using Dual-Amplification of Enzyme-Free Reaction through Universal Hybridization Chain Reaction System.

08:00 EDT 1st April 2019 | BioPortfolio

Summary of "Colorimetric Biosensor using Dual-Amplification of Enzyme-Free Reaction through Universal Hybridization Chain Reaction System."

On-site genetic detection needs to develop a sensitive and straightforward biosensor without special equipment which can detect various genetic biomarkers. Hybridization chain reaction amplifying signal isothermally could be considered as a good candidate for on-site detection. Here, we developed a novel genetic biosensor based on 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. This article is protected by copyright. All rights reserved.

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This article was published in the following journal.

Name: Biotechnology and bioengineering
ISSN: 1097-0290
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Methods used to measure the relative activity of a specific enzyme or its concentration in solution. Typically an enzyme substrate is added to a buffer solution containing enzyme and the rate of conversion of substrate to product is measured under controlled conditions. Many classical enzymatic assay methods involve the use of synthetic colorimetric substrates and measuring the reaction rates using a spectrophotometer.

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