DNA Multiway Junction Based Lateral Flow Assay for Visualized Detection of Different Types of Targets

Area of Science:

• Biotechnology
• Molecular Biology
• Assay Development

Background:

• Lateral flow assays (LFAs) offer simple, visual, and portable detection.
• Current LFAs are target-specific, requiring costly and time-consuming strip redesign for new analytes.
• This limitation hinders the broad applicability of LFA technology.

Purpose of the Study:

• To develop a universal and adaptable lateral flow assay platform.
• To overcome the target-specificity limitations of conventional lateral flow assays.
• To enable the detection of diverse biomolecules using a single assay format.

Main Methods:

• Designed a DNA multiway junction system using assistant strands (α and β) and a molecular beacon (MB).
• The junction complex bridges DNA-modified gold nanoparticles to the test zone for signal generation.
• Demonstrated detection of various targets including HBV DNA, microRNA, ATP, thrombin, and terminal deoxynucleotidyl transferase (TdT).

Main Results:

• Successfully created a DNA multiway junction that forms either a four-way or three-way junction depending on the target.
• The assay effectively linked DNA-modified gold nanoparticles for visual coloration on the test strip.
• Validated the platform's capability to detect a wide range of targets, including nucleic acids, small molecules, proteins, and enzymes.

Conclusions:

• The DNA multiway junction based lateral flow assay offers a versatile and flexible detection method.
• This approach provides a convenient and cost-effective alternative to traditional, target-specific LFAs.
• The developed platform significantly broadens the application scope of visual detection technologies.