Although lateral flow assays (LFAs) are currently being used in some point-of-care applications (POC), they cannot still be extended to a broader range of analytes for which higher sensitivities and lower detection limits are required. To overcome such drawbacks, we propose here a simple and facile alternative based on the use of delay hydrophobic barriers fabricated by wax printing so as to improve LFA sensitivity. Several wax pillar patterns were printed onto the nitrocellulose membrane in order to produce delays as well as pseudoturbulence in the microcapillary flow. The effect of the proposed wax pillar-modified devices was also mathematically simulated, corroborating the experimental results obtained for the different patterns tested afterwards for detection of HIgG as model protein in a gold nanoparticle-based LFA. The effect of the introduction of such wax-printed pillars was a sensitivity improvement of almost 3-fold compared to the sensitivity of a conventional free-barrier LFA.
Competitive LFAs are widely employed to detect targets ranging from small molecules to large proteins across diverse fields. This review explores their function using theoretical models and critically addresses factors affecting their performance.
A novel triple lines lateral-flow assay (LFA) with enhanced sensitivity for the detection of Leishmania infantum DNA in dog blood samples was designed and successfully applied. The enhanced LFA methodology takes advantage of the gold nanoparticle tags (AuNPs) conjugated to polyclonal secondary antibodies, which recognize anti-FITC antibodies. The polyclonal nature of the secondary antibodies allows for multiple binding to primary antibodies, leading to enhanced AuNP plasmonics signal. Furthermore, endogenous control consisting of the amplified dog 18S rRNA gene was introduced to avoid false negatives. Using this strategy, 0.038 spiked Leishmania parasites per DNA amplification reaction (1 parasite/100 μL of DNA sample) were detected. Detection limit of LFA was found to be lower than that of the conventional techniques. In summary, our novel LFA design is a universal and simple sensing alternative that can be extended to several other biosensing scenarios.