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Related Experiment Video

Updated: Jun 25, 2025

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Introducing Triplex Forming Oligonucleotide into Loop-Mediated Isothermal Amplification for Developing a Lateral Flow

Wei Chang1, Po-Hao Chou2,3, Cai-Tong Wu1

  • 1Master and PhD Program in Biotechnology Industry, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan.

Biosensors
|May 24, 2024
PubMed
Summary
This summary is machine-generated.

This study introduces a novel triplex-forming oligonucleotide (TFO) probe integrated into loop-mediated isothermal amplification (LAMP) to improve infectious disease detection specificity. The TFO-LAMP method enhances sequence specificity for detecting Group B Streptococcus (GBS).

Keywords:
Group B Streptococcus (GBS)Streptococcus agalactiaelateral flow assay (LFA)loop-mediated isothermal amplification (LAMP)triplex-forming oligonucleotide (TFO)

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Infectious Disease Diagnostics

Background:

  • Loop-mediated isothermal amplification (LAMP) is a sensitive method for infectious disease detection.
  • Conventional LAMP assays often lack precise sequence specificity.
  • Enhancing specificity is crucial for reliable molecular diagnostics.

Purpose of the Study:

  • To develop a more sequence-specific LAMP assay using triplex-forming oligonucleotide (TFO) probes.
  • To apply this TFO-integrated LAMP (TFO-LAMP) for the detection of Group B Streptococcus (GBS).
  • To create a nucleic acid testing biosensor based on the TFO-LAMP technology.

Main Methods:

  • Integrated a TFO probe into the LAMP process to target a specific DNA sequence (TFO targeting sequence, TTS).
  • Developed a screening method to identify high-affinity TFO sequences for LAMP integration.
  • Designed a TFO-LAMP assay utilizing a FAM-labeled TFO for detection via lateral flow biosensors.

Main Results:

  • The TFO-LAMP assay demonstrated high sequence specificity for GBS detection.
  • Sensitivity reached 85% at 300 copies and 100% at 30,000 copies of GBS.
  • The TFO-LAMP assay was successfully adapted for use with lateral flow test strips.

Conclusions:

  • Successfully developed a TFO-LAMP technology for enhanced sequence specificity in nucleic acid detection.
  • The TFO-LAMP assay is applicable for developing sensitive and specific biosensors, including lateral flow devices.
  • This technology holds potential for improved infectious disease diagnostics.