The COVID-19 pandemic has caused over 7 million deaths
worldwide
and over 1 million deaths in the US as of October 15, 2022. Virus
testing lags behind the level or availability necessary for pandemic
events like COVID-19, especially in resource-limited settings. Here,
we report a low cost, mix-and-read COVID-19 assay using a synthetic
SARS-CoV-2 sensor, imaged and processed using a smartphone. The assay
was optimized for saliva and employs 3D-printed micropipette tips
with a layer of monoclonal anti-SARS-CoV-2 inside the tip. A polymeric
sensor for SARS-CoV-2 spike (S) protein (COVRs) synthesized as a thin
film on silica nanoparticles provides 3,3′,5–5′-tetramethylbenzidine
responsive color detection using streptavidin-poly-horseradish peroxidase
(ST-poly-HRP) with 400 HRP labels per molecule. COVRs were engineered
with an NHS-PEG4-biotin coating to reduce nonspecific binding
and provide affinity for ST-poly-HRP labels. COVRs binds to S-proteins
with binding strengths and capacities much larger than salivary proteins
in 10% artificial saliva-0.01%-Triton X-100 (as virus deactivator).
A limit of detection (LOD) of 200 TCID50/mL (TCID50 = tissue culture infectious dose 50%) in artificial saliva was obtained
using the Color Grab smartphone app and verified using ImageJ. Viral
load values obtained in 10% pooled human saliva spiked with inactivated
SARS-COV-2 virus gave excellent correlation with viral loads obtained
from qPCR (p = 0.0003, r = 0.99).