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Signal-on fluorescence biosensor for microRNA-21 detection based on DNA strand displacement reaction and Mg-dependent DNAzyme cleavage. | LitMetric

AI Article Synopsis

  • MicroRNAs are important for understanding biological processes and could serve as biomarkers for diseases, highlighting the need for effective detection methods.
  • A new fluorescence assay for detecting microRNA-21 was created, utilizing DNA polymerase reactions, Mg-dependent DNAzyme reactions, and magnetic separation to enhance sensitivity.
  • This method demonstrated a low detection limit of 0.27 fM and high specificity, making it suitable for analyzing microRNA-21 in cancer patient serum, which could impact both biomedical research and clinical diagnostics.

Article Abstract

MicroRNAs have been involved into many biological processes and are regarded as disease biomarkers. Simple, rapid, sensitive and selective method for microRNA detection is crucial for early diagnosis and therapy of diseases. In this work, sensitive fluorescence assay was developed for microRNA-21 detection based on DNA polymerase induced strand displacement amplification reaction, Mg-dependent DNAzyme catalysis reaction, and magnetic separation. In the presence of target microRNA-21, amounts of trigger DNA could be produced with DNA polymerase induced strand displacement amplification reaction, and the trigger DNA could be further hybridized with signal DNA, which was labeled with biotin and AMCA dye. After introduction of Mg, trigger DNA could form DNAzyme to cleave signal DNA. After magnetic separation, the DNA fragment with AMCA dye could give fluorescence signal, which was related to microRNA-21 concentration. Based on the two efficient signal amplifications, the developed method showed high detection sensitivity with low detection limit of 0.27fM (3σ). In addition, this fluorescence strategy also possessed excellent detection specificity, and could be applied to analyze microRNA-21 expression level in serum of cancer patient. According to the obtained results, the developed fluorescence method might be a promising detection platform for microRNA-21 quantitative analysis in biomedical research and clinical diagnosis.

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Source
http://dx.doi.org/10.1016/j.bios.2017.04.049DOI Listing

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