Toward Community Surveillance: Detecting Intact SARS-CoV-2 Using Exogeneous Oligonucleotide Labels

Abstract

The persistence of the COVID-19 pandemic demands a dramatic increase in testing efficiency. Testing pooled samples for SARS-CoV-2 could meet this need; however, the sensitivity of RT-qPCR, the gold standard, significantly decreases with an increasing number of samples pooled. Here, we introduce DIVER, a method that quantifies intact virus and is robust to sample dilution. DIVER first tags viral particles with exogeneous oligonucleotides, then captures the tagged particles on ACE2-functionalized beads, and finally quantifies the oligonucleotide tags using qPCR. Using spike-presenting liposomes and Spike-pseudotyped lentivirus as SARS-CoV-2 models, we show that DIVER can detect 1x105 liposomes and 100 pfu lentivirus and can successfully identify positive samples in pooling experiments. Overall, DIVER is well-positioned for efficient sample pooling and expanded community surveillance.

DNA‐Directed Patterning for Versatile Validation and Characterization of a Lipid‐Based Nanoparticle Model of SARS‐CoV‐2

“…In validating S–liposomes, we have also developed a nanoparticle that could be used to optimize safely COVID‐19 diagnostic strategies. [ 33 ]…”

“…In validating S-liposomes, we have also developed a nanoparticle that could be used to optimize safely COVID-19 diagnostic strategies. [33] An attractive feature of our method is that it uses entirely offthe-shelf components-oligonucleotides of any 20-nucleotide sequence can be readily purchased. While we demonstrated the segregation of three different liposome types here using three oligonucleotides with different sequences, the multiplexing capability of this technique, in theory up to 4 20 oligonucleotide sequences and liposome types, makes possible the analysis of highly heterogeneous solutions of particles.…”