Cancer-Specific MicroRNA Analysis with a Nonenzymatic Nucleic Acid Circuit.
Dan ZhuBang LuYu ZhuZihao MaYaqi WeiShao SuLihua WangShiping SongYing ZhuLian-Hui WangJie ChaoPublished in: ACS applied materials & interfaces (2019)
Sensitive detection of gastric cancer-related biomarkers in human serum provides a promising means for early cancer diagnosis. Herein, we report the design of a nucleic acid circuit for gastric cancer-related microRNA-27a (miRNA-27a) detection based on dual toehold-mediated circular strand displacement amplification (CSDA). In the presence of miRNA-27a, the hybridization between miRNA-27a and probe DNA on magnetic beads through toehold 1 leads to the release of fluorescent DNA and the exposure of a new toehold 2 on linker DNA. After hybridization with catalytic DNA, CSDA is initiated and target miRNA-27a is released to participate in the next cyclic reaction; therefore, a greatly enhanced fluorescence signal is produced. The efficient magnetic separation makes the sensitive detection of miRNA-27a be accomplished within 45 min. With the efficient CSDA, the detection limit of the system (0.8 pM) is ∼100 folds lower than that of the system based on strand displacement without CSDA (79.3 pM). Furthermore, the system also showed good stability and sensitivity to discriminate single-base mismatch, which allows the detection of miRNA-27a in human serum samples. This study provides a novel platform and approach for the rapid quantitative determination of miRNA, which has great potential in clinical diagnosis and disease treatment.
Keyphrases
- nucleic acid
- sensitive detection
- loop mediated isothermal amplification
- quantum dots
- single molecule
- label free
- circulating tumor
- papillary thyroid
- air pollution
- real time pcr
- particulate matter
- molecularly imprinted
- squamous cell carcinoma
- risk assessment
- mass spectrometry
- young adults
- polycyclic aromatic hydrocarbons
- water soluble
- lymph node metastasis
- combination therapy
- human health
- electron transfer