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Development of a Lateral Flow Immunochromatographic Strip for Rapid and Quantitative Detection of Small Molecule Compounds
Published on: November 13, 2021
Second Flowing Type Lateral Flow Assay for High-Fidelity Detection.
Wenting Chang1, Siting Fu1, Wenyu Cao1
1School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100, China.
Analytical Chemistry
|May 19, 2026
Summary
This study introduces a novel two-flow lateral flow assay to prevent false positives. The second flow corrects initial results, ensuring accurate detection of targets like DNA mutations and adenosine triphosphate (ATP).
Area of Science:
- Biotechnology
- Assay Development
- Molecular Diagnostics
Background:
- Lateral flow assays offer portable, instrument-free detection.
- Target-sandwich assays are common but prone to false-positive signals.
- Existing methods struggle with high similarity analogues and non-specific interactions.
Purpose of the Study:
- To develop a high-fidelity lateral flow assay using a second flow step.
- To correct false-positive signals inherent in traditional single-flow assays.
- To enhance accuracy and reliability in target detection.
Main Methods:
- A novel second flowing-type lateral flow assay was designed.
- A second flow with displacing strands was used to validate the first flow's result.
- The assay was tested for DNA oligonucleotide mutation discrimination and adenosine triphosphate (ATP) detection.
Main Results:
- The second flow successfully corrected false-positive signals.
- Single base DNA mutations were accurately discriminated.
- Low concentrations of ATP were differentiated from background interference.
- The assay demonstrated high-fidelity detection capabilities.
Conclusions:
- The second flowing-type lateral flow assay significantly improves detection accuracy.
- This method provides a reliable solution for universally adopted target-sandwiched lateral flow detection.
- The strategy enhances the robustness of diagnostic assays, reducing misjudgments.
