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Related Experiment Video

Updated: Nov 22, 2025

Split Hybridization Probe Utilizing a DNA Fluorescent Light-up Aptamer as a Signal Reporter for Sequence-Specific Nucleic Acid Analysis

Split Hybridization Probe Utilizing a DNA Fluorescent Light-up Aptamer as a Signal Reporter for Sequence-Specific Nucleic Acid Analysis

Published on: July 8, 2025

Ligation-Enabled Fluorescence-Coding PCR for High-Dimensional Fluorescence-Based Nucleic Acid Detection.

Joon Soo Park¹, Thomas Pisanic², Ye Zhang¹

¹Department of Biomedical Engineering, Johns Hopkins University, Baltimore, Maryland 21218, United States.

Analytical Chemistry

|January 11, 2021

Summary

This summary is machine-generated.

Ligation-eNabled fluorescence-Coding PCR (LiNC PCR) enhances multiplexing for nucleic acid amplification. This novel method improves diagnostic accuracy by reducing nonspecific interactions in fluorescence-based assays.

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Area of Science:

Molecular Biology
Biotechnology
Diagnostic Assays

Background:

Polymerase chain reaction (PCR) is a cornerstone of nucleic acid amplification for diagnostics.
Traditional multiplexed PCR assays face limitations due to nonspecific interactions and fluorescence-based detection challenges.
Enhanced multiplexing is crucial for comprehensive molecular diagnostics.

Purpose of the Study:

To introduce a novel multiplexed PCR strategy, Ligation-eNabled fluorescence-Coding PCR (LiNC PCR).
To overcome technical limitations of existing fluorescence-based multiplexed PCR assays.
To significantly increase the multiplexing capacity of PCR-based diagnostic tools.

Main Methods:

Developed LiNC PCR, a method involving a preliminary ligation reaction to create unique fluorescence signatures.
Utilized universal TaqMan probes for target-specific multicolor fluorescence signal generation.
Demonstrated the technique with a two-color assay for detecting 10 ovarian cancer epigenetic biomarkers.

Main Results:

LiNC PCR exponentially enhances multiplexing capabilities compared to standard assays.
Achieved high analytical sensitivity, detecting as low as 60 template molecules.
Demonstrated no detectable target cross-talk in the developed assay.
Successfully identified 10 ovarian cancer epigenetic biomarkers.

Conclusions:

LiNC PCR offers a simple, cost-effective approach for high-dimensional multiplexing in molecular diagnostics.
The technique significantly improves the multiplexing capability of fluorescence-based PCR assays.
LiNC PCR has broad applicability in various molecular diagnostic fields.