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. 2023 Mar 15:379:133252.
doi: 10.1016/j.snb.2022.133252. Epub 2022 Dec 27.

A visual detection strategy for SARS-CoV-2 based on dual targets-triggering DNA walker

Liting Zhao  1 , Ciling Li  1 , Xinhuang Kang  1 , Yubin Li  1   2   3
Affiliations

A visual detection strategy for SARS-CoV-2 based on dual targets-triggering DNA walker

Liting Zhao et al. Sens Actuators B Chem. .

Abstract

SARS-CoV-2, a highly transmissible and mutagenic virus, made huge threats to global public health. The detection strategies, which are free from testing site requirements, and the reagents and instruments are portable, are vital for early screening and play a significant role in curbing the spread. This work proposed a silver-coated glass slide (SCGS)/DNA walker based on a dual targets-triggering mechanism, enzyme-catalyzed amplification, and smartphone data analysis, which build a portable visual detection strategy for the SARS-CoV-2 RNA-dependent RNA polymerase (RdRp) gene. By this method, the detection was reflected by the ultraviolet absorbance changes and visible color changes to the naked eye which was analyzed by Red-Green-Blue (RGB) data analysis via smartphone within 30 min, simplifying the detection process and shortening the detection time. Meanwhile, the dual targets-triggering mechanism and dual signal amplification strategy ensured detection specificity and sensitivity. Further, the practicability was verified by the detection of the real sample which provided this method an application potential in SARS-CoV-2 rapid detection.

Keywords: Biosensor; DNA Walker; SARS-CoV-2; Silver coated glass slide.

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Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

ga1
A silver-coated glass slide (SCGS)/DNA walker based on a dual targets-triggering mechanism, enzyme-catalyzed amplification, and smartphone data analysis, which build a visual detection strategy that is free from testing site requirements, and the reagents and instruments are portable, meeting the need for COVID-19 point-of-care testing (POCT).
Scheme 1
Scheme 1
Principle of the DNA walker. The substrate strand is Signal DNA - HRP, the walker strand is Capture 1/Capture 2/Target/Swing Arm/Blocker, and the DNA walker is powered by the Nb.BbvCI cleavage.
Fig. 1
Fig. 1
Feasibility assay of the strategy. (a) mixture without Target (T1 and T2), (b) mixture without T2, (c) mixture without T1, and (d) mixture with 400 pM T1 and 400 pM T2. Mixture: 5 nM CP2, 5 nM Swing Arm/Blocker, 40 U Nb.BbvCI, 120 nM H2O2 and 120 nM ABTS2-.
Fig. 2
Fig. 2
(A) The scheme of different modes. (a) Mode 1: Dual targets-triggering mechanism, (b) Mode 2: T1-triggering mechanism, (c) Mode 3: T2-triggering mechanism. (B) The heatmap shows the selectivity assay of different modes analyzed by UV–vis (a) and G values (b).
Fig. 3
Fig. 3
Analysis of the site-blocking effect in the proposed DNA walker. (A) The figures of different oligonucleotide-modified concentrations on SCGS, (a) Highly modified SCGS (200 nM), (b) Mediumly modified SCGS (60 nM), (c) loosely modified SCGS (40 nM). (B) The signal gain of different oligonucleotide-modified concentrations on SCGS.
Fig. 4
Fig. 4
(A) The absorbances of the DNA walker with different concentrations of Target, (B) Diagram of the relationship between the concentrations and the absorbances at 416 nm, (C) The G values was obtained from the smartphone. The insert shows the colorimetric results. Standard deviations were displayed by the error bars (N = 3). Experimental condition: 5 nM CP2, 5 nM Swing Arm/Blocker, 40 U Nb.BbvCI, 120 nM H2O2 and 120 nM ABTS2-.
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