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. 2021 Jan 1:171:112685.
doi: 10.1016/j.bios.2020.112685. Epub 2020 Oct 15.

One-step rapid quantification of SARS-CoV-2 virus particles via low-cost nanoplasmonic sensors in generic microplate reader and point-of-care device

Liping Huang  1 , Longfei Ding  2 , Jun Zhou  3 , Shuiliang Chen  4 , Fang Chen  4 , Chen Zhao  2 , Jianqing Xu  2 , Wenjun Hu  5 , Jiansong Ji  6 , Hao Xu  7 , Gang L Liu  8
Affiliations

One-step rapid quantification of SARS-CoV-2 virus particles via low-cost nanoplasmonic sensors in generic microplate reader and point-of-care device

Liping Huang et al. Biosens Bioelectron. .

Abstract

The spread of SARS-CoV-2 virus in the ongoing global pandemic has led to infections of millions of people and losses of many lives. The rapid, accurate and convenient SARS-CoV-2 virus detection is crucial for controlling and stopping the pandemic. Diagnosis of patients in the early stage infection are so far limited to viral nucleic acid or antigen detection in human nasopharyngeal swab or saliva samples. Here we developed a method for rapid and direct optical measurement of SARS-CoV-2 virus particles in one step nearly without any sample preparation using a spike protein specific nanoplasmonic resonance sensor. As low as 370 vp/mL were detected in one step within 15 min and the virus concentration can be quantified linearly in the range of 0 to 107 vp/mL. Measurements shown on both generic microplate reader and a handheld smartphone connected device suggest that our low-cost and rapid detection method may be adopted quickly under both regular clinical environment and resource-limited settings.

Keywords: Antibody conjugation; COVID-19; Nanoplasmonic sensor; Point of care testing; SARS-CoV-2 virus.

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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

Image 1
Graphical abstract
Fig. 1
Fig. 1
Label-free detection of SARS-CoV-2 pseudovirus with a nanoplasmonic sensor. (a) Schematic diagram of the nanoplasmonic resonance sensor for determination of SARS-CoV-2 pseudovirus concentration. (b) Photograph (Middle) of one piece of Au nanocup array chip with a drop of water on top. Scanning electron microscopy image (Left) shows the replicated nanocup array. Transmission microscopy image (Right) shows that air and water on the device surface exhibit different colors, green and far red pink, respectively.
Fig. 2
Fig. 2
Surface functionalization of nanoplasmonic sensor chip in microwell plate for detecting SARS-CoV-2 virus. (a) Integration of the nanoplasmonic sensor chip with a standard 96-well plate. (b) Schematic of nanoplasmonic sensor chip surface functionalization as well as capturing and detecting SARS-CoV-2 pseudovirus. (c, d) The typical original spectra (c) and differential spectra (d) of adjacent modification steps and detection process with 2.5 × 108 vp/mL SARS-CoV-2 pseudovirus.
Fig. 3
Fig. 3
Detection of SARS-CoV-2 pseudovirus with nanoplasmonic sensor chip by a generic microplate reader. (a) Dynamic binding curves of SARS-CoV-2 antibodies interaction with different concentrations of the SARS-CoV-2 pseudovirus over the range 0–1.6 × 1010 vp/mL at the resonant wavelength. (b) SARS-CoV-2 pseudovirus standard curve (R2 = 0.993). (c) The illustration shows the binding of spike protein on the surface of SARS-CoV-2 virus to the specific SARS-CoV-2 mAbs. SARS-CoV-2 antibodies are conjugated to the activated carboxyl groups of 11-MUA on chip surface. (d) Schematic of nanoplasmonic sensor chip detecting SARS-CoV-2 pseudovirus with AuNP enhancement. (e) ACE2 protein labeled AuNP enhanced binding curves with different concentrations of the SARS-CoV-2 pseudovirus over the range 0–6.0 × 105 vp/mL. (f) ACE2 protein labeled AuNP enhanced SARS-CoV-2 pseudovirus standard curve (R2 = 0.985). (g) SARS-CoV-2 mAbs labeled AuNP enhanced binding curves with different concentrations of the SARS-CoV-2 pseudovirus over the range 0–1.0 × 107 vp/mL. (h) SARS-CoV-2 mAbs labeled AuNP enhanced SARS-CoV-2 pseudovirus standard curve (R2 = 0.994). (i) Specificity verification test of AuNP-enhanced SARS-CoV-2 pseudovirus detection: Dynamic binding curves of SARS-CoV-2 antibodies interaction with different pseudovirus of SARS-CoV-2, SARS, MERS, and VSV at the concentration of 1.0 × 105 vp/mL.
Fig. 4
Fig. 4
Detection of SARS-CoV-2 pseudovirus with nanoplasmonic sensor chips by a point-of-care device. (a) Schematic of nanoplasmonic sensor chip cartridge detecting SARS-CoV-2 pseudovirus with a low-cost handheld point-of-care testing device. (b) The illustration shows the detection process of the sensor chip cartridge for specific SARS-CoV-2 detection. (c) Dynamic binding curves of virus and antibody interaction with different concentrations of the SARS-CoV-2 pseudovirus over the range 0–6.0 × 106 vp/mL at the resonance wavelength. (d) Specificity verification test: Dynamic binding curves of SARS-CoV-2 antibodies interaction with different pseudovirus of SARS-CoV-2, SARS, MERS, and VSV at the concentration of 5.0 × 105 vp/mL.
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