Journal
ACS NANO
Volume 15, Issue 3, Pages 3593-3611Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acsnano.0c10035
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Funding
- National Science Foundation [CBET-2029474]
- Medtronic-Bakken Endowed Chair
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LFAs, paper-based point-of-care diagnostic tools, have traditionally lower sensitivities and specificities than standard laboratory tests. Recent efforts have focused on optimizing assay kinetics, signal amplification, and sample preamplification to increase sensitivities, as well as improving specificity through high-affinity molecules and assay optimization. With continuing improvements, LFAs may soon offer performance competitive with laboratory techniques while retaining a rapid format.
Lateral flow assays (LFAs) are paper-based point-of-care (POC) diagnostic tools that are widely used because of their low cost, ease of use, and rapid format. Unfortunately, traditional commercial LFAs have significantly poorer sensitivities (mu M) and specificities than standard laboratory tests (enzyme-linked immunosorbent assay, ELISA: pM-fM; polymerase chain reaction, PCR: aM), thus limiting their impact in disease control. In this Perspective, we review the evolving efforts to increase the sensitivity and specificity of LFAs. Recent work to improve the sensitivity through assay improvement includes optimization of the assay kinetics and signal amplification by either reader systems or additional reagents. Together, these efforts have produced LFAs with ELISA-level sensitivities (pM-fM). In addition, sample preamplification can be applied to both nucleic acids (direct amplification) and other analytes (indirect amplification) prior to LFA testing, which can lead to PCR-level (aM) sensitivity. However, these amplification strategies also increase the detection time and assay complexity, which inhibits the large-scale POC use of LFAs. Perspectives to achieve future rapid (<30 min), ultrasensitive (PCR-level), and sample-to-answer POC diagnostics are also provided. In the case of LFA specificity, recent research efforts have focused on high-affinity molecules and assay optimization to reduce nonspecific binding. Furthermore, novel highly specific molecules, such as CRISPR/Cas systems, can be integrated into diagnosis with LFAs to produce not only ultrasensitive but also highly specific POC diagnostics. In summary, with continuing improvements, LFAs may soon offer performance at the POC that is competitive with laboratory techniques while retaining a rapid format.
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