Litcius/Paper detail

Hydrogel particles improve detection of SARS-CoV-2 RNA from multiple sample types

R. Barclay, Ivan Akhrymuk, Anurag Patnaik, Victoria Callahan, Caitlin W. Lehman, P. Andersen, Robbie Barbero, Stephanie M. Barksdale, Ross Dunlap, Daniel Goldfarb, Tara Jones-Roe, Ross E. A. Kelly, Bong-Chan Kim, Shida Miao, A. R. I. Munns, Denton Munns, S. Patel, E. B. Porter, R. T. Ramsey, Soumya Sibani Sahoo, Olivia Swahn, J. Warsh, Kylene Kehn‐Hall, Benjamin Lepene

2020Scientific Reports22 citationsDOIOpen Access PDF

Abstract

Here we present a rapid and versatile method for capturing and concentrating SARS-CoV-2 from contrived transport medium and saliva samples using affinity-capture magnetic hydrogel particles. We demonstrate that the method concentrates virus from 1 mL samples prior to RNA extraction, substantially improving detection of virus using real-time RT-PCR across a range of viral titers (100-1,000,000 viral copies/mL) and enabling detection of virus using the 2019 nCoV CDC EUA Kit down to 100 viral copies/mL. This method is compatible with commercially available nucleic acid extraction kits (i.e., from Qiagen) and a simple heat and detergent method that extracts viral RNA directly off the particle, allowing a sample processing time of 10 min. We furthermore tested our method in transport medium diagnostic remnant samples that previously had been tested for SARS-CoV-2, showing that our method not only correctly identified all positive samples but also substantially improved detection of the virus in low viral load samples. The average improvement in cycle threshold value across all viral titers tested was 3.1. Finally, we illustrate that our method could potentially be used to enable pooled testing, as we observed considerable improvement in the detection of SARS-CoV-2 RNA from sample volumes of up to 10 mL.

Topics & Concepts

TiterRNA extractionRNANucleic acidSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2)Viral loadVirusCoronavirus disease 2019 (COVID-19)Extraction (chemistry)VirologyChromatographyBiologyChemistryMedicineBiochemistryGeneDiseasePathologyInfectious disease (medical specialty)SARS-CoV-2 detection and testingSARS-CoV-2 and COVID-19 ResearchBiosensors and Analytical Detection