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Colorimetric biosensor using dual‐amplification of enzyme‐free reaction through universal hybridization chain reaction system
Author(s) -
Park Chae Rin,
Rhee Won Jong,
Kim Kyu Won,
Hwang Byeong Hee
Publication year - 2019
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.26978
Subject(s) - biosensor , deoxyribozyme , loop mediated isothermal amplification , detection limit , chain reaction , multiplex , horseradish peroxidase , computational biology , chemistry , microbiology and biotechnology , biology , chromatography , dna , biochemistry , enzyme , genetics , photochemistry
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.