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bioRxiv · 10.1101/2025.09.04.674023

Instrument-free quantitative detection of analytes using a novel combo lateral flow assay

Sheng, X.·Saxena, N.·xia, x.
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Abstract

Lateral flow assays (LFA) are widely regarded as the cheapest, fastest, and easiest point-of-care (PoC) tests available.1,2 A key advantage is that results can be visually interpreted without the need for specialized equipment, enabling on-site applications while keeping costs low.3 However, conventional LFAs are typically qualitative, limiting their use to simply detect the presence or absence of analytes.4 Additional limitations include narrow dynamic ranges and false negatives due to the Hook Effect in sandwich LFAs.5 In this work, we present a combo lateral flow assay (CLFA) that integrates both sandwich and competitive assay formats, enabling the quantitative detection of analytes across a broad concentration range without an external instrument. The precision of CLFA measurements can be further enhanced with artificial intelligence (AI)-based image analysis via a smartphone app. Importantly, CLFA is not subject to the Hook Effect. CLFA is easily implemented by adding just one extra test line to the standard LFA design, resulting in minimal additional manufacturing effort and cost. Given these benefits, we anticipate that CLFA will have broad applications in diagnostics and research applications. Several examples are provided to illustrate its potential.

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Sheng, X., Saxena, N., xia, x.. 2025-09-06. Instrument-free quantitative detection of analytes using a novel combo lateral flow assay. https://doi.org/10.1101/2025.09.04.674023

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