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Hydrogel particles improve detection of SARS-CoV-2 RNA from multiple sample types
R A Barclay1, I Akhrymuk2, A Patnaik1
1Ceres Nanosciences, Inc., Manassas, VA, 20110, USA.
Scientific Reports
|December 31, 2020
Summary
This study introduces a fast method using magnetic hydrogel particles to concentrate SARS-CoV-2, significantly improving viral detection in real-time RT-PCR tests. The technique enhances sensitivity for low viral loads and enables pooled testing strategies.
Area of Science:
- Biotechnology
- Virology
- Molecular Diagnostics
Background:
- Accurate and sensitive detection of SARS-CoV-2 is crucial for pandemic control.
- Current methods may struggle with low viral loads, potentially leading to false negatives.
- Efficient sample preparation is key to improving diagnostic assay performance.
Purpose of the Study:
- To develop and validate a rapid, versatile method for concentrating SARS-CoV-2 from clinical samples.
- To enhance the sensitivity of SARS-CoV-2 detection using real-time RT-PCR.
- To assess the method's compatibility with existing diagnostic workflows and its potential for pooled testing.
Main Methods:
- Utilized affinity-capture magnetic hydrogel particles for SARS-CoV-2 concentration from transport medium and saliva.
- Integrated the concentration step with RNA extraction prior to real-time RT-PCR.
- Evaluated performance using contrived samples across a range of viral titers and tested on remnant diagnostic samples.
Main Results:
- Achieved substantial improvement in SARS-CoV-2 detection across viral titers from 100-1,000,000 viral copies/mL.
- Enabled detection down to 100 viral copies/mL using the 2019 nCoV CDC EUA Kit.
- Demonstrated compatibility with commercial RNA extraction kits and a rapid heat/detergent lysis method (10-minute processing time).
- Correctly identified all positive remnant diagnostic samples and improved detection in low viral load cases (average 3.1 Ct value improvement).
- Showed potential for pooled testing, with improved SARS-CoV-2 RNA detection from sample volumes up to 10 mL.
Conclusions:
- The developed magnetic hydrogel particle method offers a rapid and effective means to concentrate SARS-CoV-2.
- This approach significantly enhances diagnostic sensitivity, particularly for low viral load samples.
- The method's versatility and compatibility support its integration into existing molecular diagnostic workflows and pooled testing strategies.
