Detecting Intact Virus Using Exogenous Oligonucleotide Labels

Autor:	Thomas R. Carey, Molly Kozminsky, Jennifer Hall, Valerie Vargas-Zapata, Kristina Geiger, Laurent Coscoy, Lydia L. Sohn
Rok vydání:	2022
Předmět:	Reverse Transcriptase Polymerase Chain Reaction SARS-CoV-2 viruses Liposomes Oligonucleotides COVID-19 Humans human activities Pandemics Sensitivity and Specificity Article Analytical Chemistry
Zdroj:	medRxiv
ISSN:	1520-6882 0003-2700
DOI:	10.1021/acs.analchem.2c00835
Popis:	The COVID-19 pandemic has revealed how an emerging pathogen can cause a sudden and dramatic increase in demand for viral testing. Testing pooled samples could meet this demand; however, the sensitivity of reverse transcription quantitative polymerase chain reaction (RT-qPCR), the gold standard, significantly decreases with an increasing number of samples pooled. Here, we introduce detection of intact virus by exogenous-nucleotide reaction (DIVER), a method that quantifies intact virus and is robust to sample dilution. As demonstrated using two models of severe acute respiratory syndrome coronavirus 2, DIVER first tags membraned particles with exogenous oligonucleotides, then captures the tagged particles on beads functionalized with a virus-specific capture agent (in this instance, angiotensin-converting enzyme 2), and finally quantifies the oligonucleotide tags using qPCR. Using spike-presenting liposomes and spike-pseudotyped lentivirus, we show that DIVER can detect 1 × 10
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::e2b35abcf8cab05c8740f35d4fef45c4 https://doi.org/10.1021/acs.analchem.2c00835 Zobrazit plný text záznamu