title: Label-free and enzyme-free colorimetric detection of microRNA by catalyzed hairpin assembly coupled with hybridization chain reaction.
abstract: In this study, a simple, label-free, and enzyme-free colorimetric biosensor has been developed for amplified detection of let-7a microRNA (miRNA) on the basis of dual signal amplification strategy. The sensing system mainly consists of four unlabeled hairpin probes termed H1, H2, H3, and H4. Upon sensing of the target miRNA, hairpin H1 is opened. Then hairpin H2 hybridizes with H1 forming H1-H2 duplex and frees the target miRNA that can be recycled to trigger another reaction cycle. In addition, the newly formed H1-H2 duplex hybridizes with hairpin H3, and this triggers the autonomous cross-opening of the two hairpins H3 and H4 through hybridization chain reaction. During this process, numerous split G-quadruplex structures are generated and further associate with cofactor hemin to form massive peroxidase-mimicking DNAzymes. The resulting DNAzymes catalyze the H2O2-mediated oxidation of colorless 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS(2-)) to the green-colored ABTS(•-), inducing a remarkably amplified colorimetric signal. This newly developed sensing system exhibits high sensitivity toward miRNA with a detection limit of 7.4fM and a large dynamic range of 6 orders of magnitude from 10fM to 10nM. Furthermore, it exhibits a good performance to discriminate single-base difference among the miRNA family members and holds a great potential for early diagnosis in gene-related diseases.
authors.affiliation: Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, Jiangsu 214063, China. Electronic address: wuhao@jsinm.org.
authors.forename: Hao
authors.initials: H
authors.lastname: Wu
authors.affiliation: Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, Jiangsu 214063, China.
authors.forename: Yaling
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, Jiangsu 214063, China.
authors.forename: Hongyong
authors.initials: H
authors.lastname: Wang
authors.affiliation: Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, Jiangsu 214063, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Wu
authors.affiliation: Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, Jiangsu 214063, China.
authors.forename: Feifan
authors.initials: F
authors.lastname: Zhu
authors.affiliation: Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, Jiangsu 214063, China. Electronic address: zoupei@jsinm.org.
authors.forename: Pei
authors.initials: P
authors.lastname: Zou
mesh_terms: D052160:Benzothiazoles / Q000737:chemistry; D015374:Biosensing Techniques / Q000379:methods*; D003124:Colorimetry / Q000379:methods; D004396:Coloring Agents / Q000737:chemistry; D021881:DNA, Catalytic / Q000737:chemistry; D054856:G-Quadruplexes; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry; D057230:Limit of Detection; D008168:Lung / Q000737:chemistry; D035683:MicroRNAs / Q000032:analysis*; D009693:Nucleic Acid Hybridization / Q000379:methods; D013451:Sulfonic Acids / Q000737:chemistry
chemical_list: D052160:Benzothiazoles; D004396:Coloring Agents; D021881:DNA, Catalytic; D035683:MicroRNAs; D013451:Sulfonic Acids; C487931:mirnlet7 microRNA, human; C002502:2,2'-azino-di-(3-ethylbenzothiazoline)-6-sulfonic acid; D006861:Hydrogen Peroxide
keywords: Colorimetric biosensor; DNAzyme; Dual signal amplification; Split G-quadruplex structure; miRNA