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Updated: Sep 26, 2025

Open-Source Miniature Fluorimeter to Monitor Real-Time Isothermal Nucleic Acid Amplification Reactions in Resource-Limited Settings

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Published on: February 3, 2021

Ligation-free isothermal nucleic acid amplification.

Jeong Moon¹, Jayeon Song², Hyowon Jang²

¹Bionanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon, 34141, Republic of Korea; Department of Chemical and Biomolecular Engineering (BK21+ Program), Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon, 34141, Republic of Korea.

Biosensors & Bioelectronics

|April 17, 2022

Summary

This study introduces a novel ligation-free DNA extension method for isothermal nucleic acid amplification. The technique enables sensitive detection of target RNA, showing promise for cancer companion diagnostics and various gene detection platforms.

Keywords:

Cancer diagnosis Isothermal amplification Ligation-free Nucleic acid Phosphorothioate probe

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Visual Detection of Multiple Nucleic Acids in a Capillary Array

Visual Detection of Multiple Nucleic Acids in a Capillary Array

Published on: November 15, 2017

Area of Science:

Molecular Biology
Biotechnology
Diagnostics

Background:

Nucleic acid amplification is crucial for sensitive molecular detection.
Existing methods often require ligation steps, adding complexity and time.
Developing ligation-free amplification is essential for simpler, faster diagnostics.

Purpose of the Study:

To develop a novel ligation-free DNA extension method for nucleic acid amplification.
To enable sensitive detection of target RNA using isothermal amplification.
To explore the application of this method in cancer companion diagnostics.

Main Methods:

A ligation-free DNA extension reaction involving two fragmented probes hybridized to target RNA.
DNA elongation occurs from a forward probe to a phosphorothioated-hairpin probe.
Simultaneous DNA elongation at a nick site and self-priming region amplifies CRISPR/Cas12a binding sites.

Main Results:

Achieved a sensitivity of 49.2 fM for target RNA detection.
Demonstrated feasibility for detecting two types of mRNA.
Validated the method using mRNA from human cells, mouse tissues, and urine samples.

Conclusions:

The developed ligation-free isothermal nucleic acid amplification system offers high sensitivity and broad applicability.
This method holds significant potential for cancer companion diagnostics.
The system is expected to be widely adopted in various gene detection platforms.