期刊
SENSORS AND ACTUATORS B-CHEMICAL
卷 363, 期 -, 页码 -出版社
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2022.131858
关键词
Y shape probe; Ternary DNA nanomachine; MiRNA determination; Isothermal all -in amplification; Fluorescent sensing
资金
- National Natural Science Foundation of China [21804028]
- Key Research and Development Program of Anhui Province [202004h07020029, 202104a07020008]
- Natural Science Foundation of Anhui Province [1908085QC121, 2108085MB65]
- Natural Science Research Project for Universities from the Education Depart-ment of Anhui Province in China [KJ2020A0522, KJ2019A0513, KJ2019ZD37]
- Natural Science Project of Fuyang Normal University [2020FSKJ06ZD]
- Scientific Research and Innovation Team of Fuyang Normal University [KYTD202001]
Researchers proposed a ternary DNA nanomachine for microRNA determination, which can quantitatively detect miRNA as low as 1 fM with high sensitivity and specificity.
The construction of DNA nanodevices is vital for disease diagnosis and biomedical study. Herein, we proposed a ternary DNA nanomachine for microRNA determination using a molecular beacon (MB) coupled Y shape probe (MB-YP). The MB-YP was a tripartite junction scaffold connected with MB-Y-I, Y-II, and Y-III. The target miRNA firstly recognizes MB-Y-I, activating the MB-Y-I-DNA nanomachine to produce endogenous primer I (EPI) responsible for the activation of Y-II-DNA nanomachine. The Y-II-DNA nanomachine then allows the production of numerous EPII, which in turn activates the Y-III-DNA nanomachine to produce EPIII for re-activating MB-Y-I-DNA nanomachine. The stepwise operated MB-Y-I-DNA nanomachine, Y-II-DNA nanomachine, and Y-III-DNA nanoma-chine constituted the ternary DNA nanomachine and the repeated use of EPI, EPII, and EPIII permits it an all-in amplification. These features lead to the opening of MB-YI for emitting strong fluorescence. As low as 1 fM miRNA can thus be quantitatively detected and the linear response of the miRNA is from 1 fM to 100 pM (six orders of magnitude). Moreover, the nanomachine is specific to distinguish target miRNA from one-base mutant, inserted, and deleted target analogs. Its practicability was demonstrated by confirming miRNA-21 is overex-pressed in clinical human serums from lung cancer patients than healthy donors.
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