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Related Concept Videos

Labeling DNA Probes03:31

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...

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Real-time Imaging of Single Engineered RNA Transcripts in Living Cells Using Ratiometric Bimolecular Beacons
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Light-up RNA aptamer-based T-NASBA: a one-pot, label-free strategy for miRNA detection.

Yao Fu1, Xuejuan Pei1, Junhua Cao2

  • 1Department of Forensic Medicine, Chongqing Medical University, Chongqing, 400016, China.

Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry
|August 5, 2025
PubMed
Summary

We created a simple, cost-effective method for detecting microRNAs (miRNAs) without labels or complex equipment. This new platform offers accurate and sensitive miRNA detection, ideal for point-of-care diagnostics and research.

Keywords:
Fluorescent RNA aptamersIsothermal amplificationMicroRNANASBAToehold-mediated strand displacement

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Area of Science:

  • Biotechnology
  • Molecular Diagnostics
  • Cancer Research

Background:

  • MicroRNAs (miRNAs) are crucial regulators of cellular functions.
  • miRNA dysregulation is associated with various diseases, especially cancer.
  • Existing miRNA detection methods are often costly and complex.

Purpose of the Study:

  • To develop a novel, label-free, and cost-effective platform for microRNA detection.
  • To integrate toehold-mediated strand displacement (TMSD) with nucleic acid sequence-based amplification (NASBA) for enhanced sensitivity.
  • To provide a simplified alternative for miRNA analysis without specialized instrumentation.

Main Methods:

  • A one-pot platform combining TMSD with NASBA was developed.
  • A TMSD-regulated hairpin primer triggers NASBA upon target miRNA binding.
  • NASBA generates a light-Up RNA aptamer for real-time fluorescence detection.

Main Results:

  • The platform achieved a low detection limit of 4.31 pM for miRNAs.
  • A strong linear correlation was observed between fluorescence and miRNA concentration (10 pM to 1 nM).
  • High precision and reproducibility were confirmed in human serum samples (99.85%–108.28% recovery).

Conclusions:

  • The developed T-NASBA platform offers a sensitive, cost-effective, and label-free approach for miRNA detection.
  • This method eliminates the need for complex equipment and fluorescent labels.
  • T-NASBA is suitable for point-of-care diagnostics and various research applications in miRNA analysis.