| Autor: |
Kylene Kehn-Hall, Caitlin W. Lehman, Ross Kelly, Ben Lepene, Robbie Barbero, Olivia Swahn, Robert A Barclay, Rosalind Ramsey, Shida Miao, Patrick Andersen, Denton Munns, Ivan V. Akhrymuk, Erica Porter, Amy Munns, Daniel Goldfarb, Brianna Kim, Stephanie Barksdale, Samip Patel, Saswata Sahoo, Jack Warsh, Ross Dunlap, Victoria Callahan, Tara Jones-Roe, Anurag Patnaik |
| Rok vydání: |
2020 |
| Předmět: |
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| Popis: |
Here we present a rapid and versatile method for capturing and concentrating SARS-CoV-2 from transport medium and saliva using affinity-capture magnetic hydrogel particles. We demonstrate that the method concentrates virus prior to RNA extraction, thus significantly improving detection of the virus using a real-time RT-PCR assay across a range of viral titers, from 100 to 1,000,000 viral copies/mL; in particular, detection of virus in low viral load samples is enhanced when using the method coupled with the IDT 2019-nCoV CDC EUA Kit. This method is compatible with commercially available nucleic acid extraction kits, as well with a simple heat and detergent method. Using transport medium diagnostic remnant samples that previously had been tested for SARS-CoV-2 using either the Abbott RealTime SARS-CoV-2 EUA Test (n=14) or the Cepheid Xpert Xpress SARS-CoV-2 EUA Test (n=35), we demonstrate that our method not only correctly identifies all positive samples (n = 17) but also significantly improves detection of the virus in low viral load samples. The average improvement in cycle threshold (Ct) value as measured with the IDT 2019-nCoV CDC EUA Kit was 3.1; n = 10. Finally, to demonstrate that the method could potentially be used to enable pooled testing, we spiked infectious virus or a confirmed positive diagnostic remnant sample into 5 mL and 10 mL of negative transport medium and observed significant improvement in the detection of the virus from those larger sample volumes. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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