期刊
SENSORS AND ACTUATORS B-CHEMICAL
卷 344, 期 -, 页码 -出版社
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2021.130280
关键词
Electrochemical sensing; Isothermal amplification; Digestion-to-growth regulation microRNA detection; Signal enhancement
资金
- Anhui Key RD Project [201904d07020016]
- Ministry of Science and Technology of China [2018YFC1603606, 2017YFF0208600]
- NSFC [21804028]
- China Agriculture Research System48 [CARS48]
- Anhui Provincial Natural Science Foundation [1908085QC121]
- China Postdoctoral Science Foundation [225351]
- Funda-mental Research Fund for Central University [JZ2019HGTB0068, PA2017GDQT0018, EWPL201905]
A DG-TIS-based electrochemical biosensor was developed for the ultrasensitive detection of miRNA-21, showing wide linear response range, low detection limit, and excellent specificity. The biosensor could effectively interrogate the expression levels of miRNA-21 in cancer cells and spiked human serum, making it an ideal analytical tool for diagnostic applications.
Ultrasensitive detection of microRNA (miRNA) has well-demonstrated values for cancer diagnosis, prognosis and therapy. In this work, taking advantages of the designed versatility of oligonucleotides and the robustness of electrochemistry, a unique digestion-to-growth regulated tandem isothermal amplification (DG-TIS) based electrochemical biosensor has been developed for miRNA-21. In DG-TIS system, two hairpin probes (HP1 and HP2) are used as the enzymatic reaction unit. Target miRNA firstly hybridize with HP1 to induce the T7 exonuclease aided-digestion amplification (T7 Exo-ADA) for HP1 digestion. The produced hydrolysis residue strand then captures HP2 to induce the Klenow polymerase promoted-growth amplification (Klenow Poly-PGA) for HP2 growth. The tandem connection of T7 Exo-ADA and Klenow Poly-PGA makes the DG-TIS finally contribute considerably to the electrochemical responses by stacking tremendous electrochemical active tags. This biosensor with the unique digestion-to-growth regulation for miRNA sensing, as a proof of concept, showed a wide linear response range over 4 orders of magnitude, a lower detection limit down to 34.8 fM, and an excellent specificity to easily distinguish even one-base mismatched analyte. Impressively, this electrochemical biosensor also offered an acceptable practical applicability to interrogate the expression levels of miRNA-21 in cancer cells and spiked human serum. These features do make DG-TIS based electrochemical biosensor an ideal analytical tool for diagnostic applications.
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