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URL: https://pubmed.ncbi.nlm.nih.gov/32743622/

⇱ A modular microarray imaging system for highly specific COVID-19 antibody testing - PubMed

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Abstract

To detect the presence of antibodies in blood against SARS-CoV-2 in a highly sensitive and specific manner, here we describe a robust, inexpensive ($200), 3D-printable portable imaging platform (TinyArray imager) that can be deployed immediately in areas with minimal infrastructure to read coronavirus antigen microarrays (CoVAMs) that contain a panel of antigens from SARS-CoV-2, SARS-1, MERS, and other respiratory viruses. Application includes basic laboratories and makeshift field clinics where a few drops of blood from a finger prick could be rapidly tested in parallel for the presence of antibodies to SARS-CoV-2 with a test turnaround time of only 2-4 h. To evaluate our imaging device, we probed and imaged coronavirus microarrays with COVID-19-positive and negative sera and achieved a performance on par with a commercial microarray reader 100× more expensive than our imaging device. This work will enable large scale serosurveillance, which can play an important role in the months and years to come to implement efficient containment and mitigation measures, as well as help develop therapeutics and vaccines to treat and prevent the spread of COVID-19.

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Conflict of interest statement

The coronavirus antigen microarray is intellectual property of the Regents of the University of California that is licensed for commercialization to Nanommune Inc. (Irvine, CA), a private company for which Philip L. Felgner is the largest shareholder and several co-authors (de Assis, Jain, Nakajima, Jasinskas, and Khan) also own shares. Nanommune Inc. has a business partnership with Sino Biological Inc. (Beijing, China) which expressed and purified the antigens used in this study. Weian Zhao is the founder of Velox Biosystems Inc. (Irvine, CA) that develops diagnostics including for COVID-19.

Figures

👁 Fig. 1
Fig. 1. TinyArray imager design. (a) Workflow: after probing of the antigen microarrays, images are taken where the fluorescence intensities corresponding to the relative antibody concentration are quantified. (b) The microarray was LED-illuminated (470 nm) from the top and imaged through long pass and band pass filters with an OmniVision OV5647 sensor module. Illumination was controlled and images were acquired with a single board computer (Raspberry Pi 4). (c) CAD design of the microarray imager. (d) 3D printed and assembled prototype together with a Raspberry Pi 4 single board computer interfacing the camera and 75 mm × 25 mm × 1 mm microarray slide inserted into the device. Scale bars, 30 mm.
👁 Fig. 2
Fig. 2. Fluorescence images and quantification of printed microarrays with 280 μm spaced, 150 μm-diameter dots of labeled with quantum dot probes. (a and b) Fluorescence images of 2 × 4 microarray pads (7 mm × 7 mm each) taken with the commercial ArrayCam 400-S and the TinyArray imager, respectively. Fluorescence intensity is represented on a pseudo rainbow color scale. (c) Layout and relative concentrations of the serial dilutions of QD655–streptavidin microarray dots imaged in panels (a and b). (d and e) Quantitative analysis of the background-subtracted median intensities in the top (panel d) and bottom QD655–streptavidin dot rows (panel e) of the images taken with the ArrayCam (left column) and the TinyArray imager (right column) as shown in panels (a and b). (f) Raw image of four microarray pads probed with human serum samples and developed with secondary antibodies conjugated to QD655. (g) Background-subtracted microarray image in pseudo rainbow color scale. (h) Graph and linear regression (R2 > 0.85) of the background-subtracted median fluorescence intensities of the same microarray sample as obtained with the TinyArray imager prototype and the ArrayCam 400-S. Scale bars, 2 mm.
👁 Fig. 3
Fig. 3. Fluorescence images and data analysis of CoVAM probed with positive sera and stained for human IgG. (a) Four exemplary fluorescence images acquired with the Array Cam 400-S. (b) Corresponding TinyArray images. (c) Background-subtracted median fluorescence intensities obtained for each microarray spot with the Array Cam 400-S and the TinyArray imager that were normalized and plotted against each other; linear regressions were performed and R2 values were calculated. (d) Heat maps of 9 SARS-CoV-2-positive and 10 negative control samples generated from the Array Cam 400-S (top row) and TinyArray imager data (bottom row). Gray/black/red colors indicate low/medium/high antibody prevalence. (e and f) Statistical analysis of the seven SARS-CoV-2 antigens in the CoVAM for positive sera for the ArrayCam and the TinyArray imager data. Scale bar, 2 mm.

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