Development of nucleic acid based lateral flow assays for SARS-CoV-2 detection

Derin, Dilek Çam; Gültekin, Enes; Taşkın, Irmak İçen; Yakupoğulları, Yusuf

doi:10.1016/j.jbiosc.2022.11.001

Cited by 5 publications

(3 citation statements)

References 28 publications

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“…Aggregates of cysteamine-AuNPs/streptavidin-AuNPs were used as nano-amplifiers of colorimetric/SERS LFA signals formed in the presence of extracted target SARS-CoV-2 RNA from 30 positive nasal/nasopharyngeal swab samples (30 min test) and two specific 6-carboxyfluorescein (FAM)/biotin-labeled antisense DNA oligonucleotide probes (20 bp). The anti-FAM antibody and anti-streptavidin antibody were used for the T-line and C-line, respectively [129].…”

Section: Nucleic Acid Lfas: Fourth Generationmentioning

confidence: 99%

Updates on the Biofunctionalization of Gold Nanoparticles for the Rapid and Sensitive Multiplatform Diagnosis of SARS-CoV-2 Virus and Its Proteins: From Computational Models to Validation in Human Samples

Ionescu

2023

IJMS

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Since the outbreak of the pandemic respiratory virus SARS-CoV-2 (COVID-19), academic communities and governments/private companies have used several detection techniques based on gold nanoparticles (AuNPs). In this emergency context, colloidal AuNPs are highly valuable easy-to-synthesize biocompatible materials that can be used for different functionalization strategies and rapid viral immunodiagnosis. In this review, the latest multidisciplinary developments in the bioconjugation of AuNPs for the detection of SARS-CoV-2 virus and its proteins in (spiked) real samples are discussed for the first time, with reference to the optimal parameters provided by three approaches: one theoretical, via computational prediction, and two experimental, using dry and wet chemistry based on single/multistep protocols. Overall, to achieve high specificity and low detection limits for the target viral biomolecules, optimal running buffers for bioreagent dilutions and nanostructure washes should be validated before conducting optical, electrochemical, and acoustic biosensing investigations. Indeed, there is plenty of room for improvement in using gold nanomaterials as stable platforms for ultrasensitive and simultaneous “in vitro” detection by the untrained public of the whole SARS-CoV-2 virus, its proteins, and specific developed IgA/IgM/IgG antibodies (Ab) in bodily fluids. Hence, the lateral flow assay (LFA) approach is a quick and judicious solution to combating the pandemic. In this context, the author classifies LFAs according to four generations to guide readers in the future development of multifunctional biosensing platforms. Undoubtedly, the LFA kit market will continue to improve, adapting researchers’ multidetection platforms for smartphones with easy-to-analyze results, and establishing user-friendly tools for more effective preventive and medical treatments.

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Section: Nucleic Acid Lfas: Fourth Generationmentioning

confidence: 99%

Updates on the Biofunctionalization of Gold Nanoparticles for the Rapid and Sensitive Multiplatform Diagnosis of SARS-CoV-2 Virus and Its Proteins: From Computational Models to Validation in Human Samples

Ionescu

2023

IJMS

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“…There is also a need for intricate molecular assemblies, often with various pairs of antibody-antigen, which introduces growing entropy, as the number of test lines increases [9]. Furthermore, the production of ssDNA products that could hybridize with oligonucleotide probes attached on the strip often demand a post-amplification processing step (by using CRISPR, or specific enzymatic treatments) that adds time and optimization needs to the assay [8,9,12].…”

Section: Introductionmentioning

confidence: 99%

The potential of tailed amplicons for SARS-CoV-2 detection in Nucleic Acid Lateral Flow Assays

Vindeirinho,

Oliveira,

Pinho

et al. 2024

PLoS ONE

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Nucleic Acid Lateral Flow Assays (NALFAs) are a promising solution for the point-of-care detection of viruses like SARS-CoV-2. However, they show some drawbacks, such as the great dependency on the use of antibodies and the need for post-amplification protocols that enable the preparation of amplicons for effective readings, as well as low sensitivity. Here, we developed amplicons of a specific SARS-CoV-2 gene tailed with single-strand DNA (ssDNA) sequences to hybridize with DNA probes immobilized on the NALFA strips, thus overcoming the aforementioned problems. Results have shown that tailed primers have not compromised the amplification efficiency and allowed the correct detection of the amplicons in the lateral flow strip. This approach has presented a limit of detection (LOD) of 25 RNA copies /reaction mix (1 copy/μL) and the test of cross-reactivity with other related viruses has not shown any cross-reactivity. Twenty clinical samples were evaluated by NALFA and simultaneously compared with the gold standard RT-qPCR protocol, originating equal results. Although the number of clinical specimens tested being relatively small, this indicates a sensitivity and specificity both of 100%. In short, an alternative NALFA was successfully implemented, rendering an accurate route for SARS-CoV-2 diagnosis, compatible with low-resource settings.

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“…To facilitate the easy differentiation of the results, various detection methods are used, either visually or indirectly through instruments such as portable fluorescence detectors. Examples of detection methods include real-time fluorescence detection [73], lateral flow detection [74][75][76], visual inspection [77,78], and CRISPR/Cas detection [79,80]. Fluorescence detection involves exposing the fluorescent material in the system to a specific wavelength of light, causing it to emit fluorescence.…”

Section: Introductionmentioning

confidence: 99%

Advances in Simple, Rapid, and Contamination-Free Instantaneous Nucleic Acid Devices for Pathogen Detection

Wang,

Zhou

et al. 2023

Biosensors

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Pathogenic pathogens invade the human body through various pathways, causing damage to host cells, tissues, and their functions, ultimately leading to the development of diseases and posing a threat to human health. The rapid and accurate detection of pathogenic pathogens in humans is crucial and pressing. Nucleic acid detection offers advantages such as higher sensitivity, accuracy, and specificity compared to antibody and antigen detection methods. However, conventional nucleic acid testing is time-consuming, labor-intensive, and requires sophisticated equipment and specialized medical personnel. Therefore, this review focuses on advanced nucleic acid testing systems that aim to address the issues of testing time, portability, degree of automation, and cross-contamination. These systems include extraction-free rapid nucleic acid testing, fully automated extraction, amplification, and detection, as well as fully enclosed testing and commercial nucleic acid testing equipment. Additionally, the biochemical methods used for extraction, amplification, and detection in nucleic acid testing are briefly described. We hope that this review will inspire further research and the development of more suitable extraction-free reagents and fully automated testing devices for rapid, point-of-care diagnostics.

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Development of nucleic acid based lateral flow assays for SARS-CoV-2 detection

Cited by 5 publications

References 28 publications

Updates on the Biofunctionalization of Gold Nanoparticles for the Rapid and Sensitive Multiplatform Diagnosis of SARS-CoV-2 Virus and Its Proteins: From Computational Models to Validation in Human Samples

Updates on the Biofunctionalization of Gold Nanoparticles for the Rapid and Sensitive Multiplatform Diagnosis of SARS-CoV-2 Virus and Its Proteins: From Computational Models to Validation in Human Samples

The potential of tailed amplicons for SARS-CoV-2 detection in Nucleic Acid Lateral Flow Assays

Advances in Simple, Rapid, and Contamination-Free Instantaneous Nucleic Acid Devices for Pathogen Detection

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