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Related Experiment Video

Updated: Jan 6, 2026

Highly Efficient Ligation of Small RNA Molecules for MicroRNA Quantitation by High-Throughput Sequencing
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λ-exonuclease-driven split G-quadruplex hybridization-based DNA walking system for sensitive microRNA quantification.

Huijing Li1, Miaohua Ruan2

  • 1Department of Pediatrics, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325000, Zhejiang, China.

Biotechnology Letters
|October 9, 2025
PubMed
Summary

A new fluorescence biosensor accurately detects microRNAs (miRNAs) for newborn pneumonia diagnosis. This method uses a DNA walker and split G-quadruplex for sensitive, label-free detection in clinical samples.

Keywords:
DNA walkerG-quadruplexMicroRNAPneumoniaThT (thioflavin T)

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Area of Science:

  • Biotechnology
  • Molecular Diagnostics
  • Biochemistry

Background:

  • Accurate microRNA (miRNA) measurement is crucial for diagnosing newborn pneumonia.
  • Existing detection methods may lack sensitivity or require complex procedures.

Purpose of the Study:

  • To develop a simple, sensitive, and accurate fluorescence-based biosensor for miRNA detection.
  • To enable label-free identification of miRNAs using a novel DNA walker and signal amplification system.

Main Methods:

  • Utilized a lambda-exonuclease (λ-Exo)-driven DNA walker system.
  • Employed split G-quadruplex (split-G4) structures for signal amplification.
  • Detected the reformed G-quadruplex structure using the fluorescent dye thioflavin T (ThT).

Main Results:

  • Achieved highly sensitive and label-free miRNA identification.
  • Demonstrated greatly elevated signal amplification efficiency due to the DNA walking mechanism.
  • Exhibited low background noise and good dependability.
  • Successfully applied the technique to clinical specimens for disease diagnosis.

Conclusions:

  • The developed biosensor offers a robust platform for sensitive miRNA quantification.
  • This technique shows significant potential for the early diagnosis of newborn pneumonia.
  • The simplicity and accuracy make it suitable for clinical applications.