Molecularly Imprinted Polymers as Biomimetic Test Zones in Paper-Based Nucleic Acid Assays-Comparing Vertical and...

Area of Science:

• Biomaterials Science
• Analytical Chemistry
• Molecular Diagnostics

Background:

• Point-of-care nucleic acid testing requires reliable synthetic recognition elements for enhanced sensitivity and speed.
• Molecularly imprinted polymers (MIPs) offer a stable, biomimetic alternative to traditional biological receptors.

Purpose of the Study:

• To synthesize and optimize a biotin-specific molecularly imprinted polymer (MIP).
• To integrate the biotin-MIP into paper-based nucleic acid vertical flow (NAVF) and lateral flow (NALF) assay formats.
• To evaluate the performance of NAVF and NALF assays for detecting double-tagged PCR amplicons from Escherichia coli.

Main Methods:

• Synthesis of a biotin-specific MIP using an optimized protocol.
• Integration of the MIP as a test zone in paper-based NAVF and NALF devices.
• Detection of double-tagged PCR amplicons from Escherichia coli using both NAVF and NALF platforms.

Main Results:

• The NAVF assay achieved a visual readout in 3 minutes with a limit of detection (LOD) of 1.00 × 10^-2 ng mL^-1.
• The NALF assay completed in under 15 minutes, yielding a visual LOD of 3.17 × 10^-2 ng mL^-1.
• Both platforms demonstrated the effectiveness of biotin-MIPs for rapid nucleic acid detection.

Conclusions:

• Biotin-MIPs are versatile and stable synthetic receptors suitable for rapid, low-cost paper-based nucleic acid assays.
• The NAVF format excels in speed and design flexibility for point-of-care applications.
• The NALF format offers higher analytical sensitivity, making it suitable for applications requiring greater precision.