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Developing a Modular Platform for the Detection of microRNAs Using Rolling Circle Amplification and Multi-Primed

Mahboobeh Nasiri1, Kyle Nowlin1, Reza Zadegan1

  • 1Department of Nanoengineering, Joint School of Nanoscience & Nanoengineering, North Carolina Agricultural and Technical State University, Greensboro, North Carolina, USA.

Journal of Molecular Recognition : JMR
|November 17, 2025
PubMed
Summary
This summary is machine-generated.

We developed a sensitive fluorescence biosensor using rolling circle amplification (RCA) and multi-primed chain amplification (MCA) to detect low concentrations of microRNAs (miRNAs). This modular platform offers robust and adaptable molecular diagnostics.

Keywords:
biosensorcancermicroRNAmulti‐primed chain amplificationrolling circle amplification

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Biotechnology

Background:

  • MicroRNAs (miRNAs) are crucial regulators in biological processes and disease biomarkers.
  • Low miRNA concentrations in samples challenge accurate detection and quantification.
  • Sensitive detection methods are vital for molecular diagnostics and biomarker discovery.

Purpose of the Study:

  • To develop a highly sensitive fluorescence-based biosensing platform for microRNA detection.
  • To overcome the challenge of low microRNA concentrations in biological samples.
  • To create a modular and reconfigurable system for detecting various microRNAs.

Main Methods:

  • Integrated rolling circle amplification (RCA) and multi-primed chain amplification (MCA) for signal enhancement.
  • Designed a modular system allowing flexible reconfiguration for different miRNA targets.
  • Utilized a universal initiator (miR-i) and hybridization with specific miRNAs (miR-A, miR-D) for amplification.
  • Employed molecular beacons and spectrofluorometric analysis for fluorescence detection.

Main Results:

  • Achieved a strong linear correlation between miRNA concentration and fluorescence intensity.
  • Demonstrated a limit of detection (LOD) and limit of quantification (LOQ) below 10 pM in buffer and human serum.
  • Confirmed the platform's high sensitivity, robustness, and reliability in complex biological matrices.
  • Validated the modular design for adaptable miRNA detection.

Conclusions:

  • The developed biosensing platform significantly enhances sensitivity for microRNA detection.
  • Its modular architecture allows for broad applicability in detecting diverse non-coding RNAs.
  • This technology holds promise as a versatile tool for molecular diagnostics across various conditions.
  • The platform addresses the critical need for sensitive and reliable detection of low-abundance biomarkers.