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Real Time RT-PCR

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...

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Updated: Jan 8, 2026

Detection of Rare Mutations in CtDNA Using Next Generation Sequencing

Detection of Rare Mutations in CtDNA Using Next Generation Sequencing

Published on: August 24, 2017

MED: Multiplexed, Entropy-Driven Catalysis, Single-Nucleotide-Resolved ctDNA Detection.

Lijia Yuan^1,2, Ting Xiao¹, Jing Li¹

¹Department of Clinical Laboratory Medicine, Southwest Hospital, Third Military Medical University, 30 Gaotanyan, Shapingba, Chongqing 400038, China.

Analytical Chemistry

|December 18, 2025

Summary

This summary is machine-generated.

A novel multiplexed assay, MED, enables enzyme-free, single-nucleotide-resolved detection of circulating tumor DNA (ctDNA) at the point-of-care. This technology offers rapid, accessible results for enhanced cancer diagnosis and monitoring.

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Last Updated: Jan 8, 2026

Detection of Rare Mutations in CtDNA Using Next Generation Sequencing

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Visual Detection of Multiple Nucleic Acids in a Capillary Array

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Published on: November 15, 2017

Area of Science:

Biotechnology
Molecular Diagnostics
Nanotechnology

Background:

Real-time monitoring of circulating tumor DNA (ctDNA) is crucial for early cancer detection, treatment response assessment, and identifying resistance.
Existing ctDNA detection methods often lack the scalability and accessibility required for point-of-care applications.
There is a significant need for sensitive assays capable of resolving single-nucleotide variations in ctDNA.

Purpose of the Study:

To develop a multiplexed, single-nucleotide-resolved assay for the detection of ctDNA.
To enable enzyme-free nucleic acid amplification and visual detection of ctDNA at the point-of-care.
To assess the diagnostic accuracy and clinical potential of the developed assay.

Main Methods:

Introduction of a multiplexed, entropy-driven catalysis (EDC) assay for single-nucleotide-resolved ctDNA detection.
Utilizing a triple amplification strategy involving EDC, silver nanoparticle (AgNP) catalyzed Ag+ release, and urease-catalyzed urea hydrolysis for colorimetric readout.
Integration with bioactive paper and smartphone readout for point-of-care application.

Main Results:

The MED assay demonstrated 100% concordance with sequencing for detecting a panel of four ctDNAs in 40 blood specimens.
The assay achieved over 85% accuracy in distinguishing between cancer and non-cancer samples.
Integrated point-of-care format provided rapid (within 45 min) and accessible results with over 70% accuracy.

Conclusions:

The MED assay offers a scalable, enzyme-free, and single-nucleotide-resolved method for ctDNA detection.
The point-of-care format provides rapid, multiplexed, and accessible results with significant clinical potential.
This technology can enhance early cancer diagnosis, dynamic monitoring, and personalized therapeutic strategies.