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Rapid SARS-CoV-2 testing in primary material based on a novel multiplex RT-LAMP assay
Bernhard Schermer1,2,3, Francesca Fabretti1,2,3, Maximilian Damagnez4
1Department II of Internal Medicine, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany.
Plos One
|November 2, 2020
Summary
Novel multiplexed reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays offer a sensitive and rapid point-of-care solution for SARS-CoV-2 detection. This PCR-independent method is ideal for widespread pandemic testing.
Area of Science:
- Molecular Biology
- Virology
- Public Health
Background:
- Effective management of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) outbreaks necessitates rapid, large-scale population testing.
- Current point-of-care (POC) testing faces challenges due to the need for thermal cycling and nucleic acid isolation in standard PCR methods.
Purpose of the Study:
- To develop and validate PCR-independent methods for rapid SARS-CoV-2 RNA detection at the point-of-care.
- To enhance the sensitivity of isothermal amplification methods for reliable diagnostics.
Main Methods:
- Evaluated PCR-independent methods, including reverse transcription loop-mediated isothermal amplification (RT-LAMP) and specific high-sensitivity enzymatic reporter unlocking (SHERLOCK).
- Developed and optimized a novel multiplexed RT-LAMP assay for increased sensitivity using primary nasopharyngeal swab samples.
- Directly compared and combined the novel RT-LAMP assays with SHERLOCK.
Main Results:
- Standard RT-LAMP assays showed satisfactory specificity but lacked the sensitivity of quantitative real-time polymerase chain reaction (qPCR).
- The novel multiplexed RT-LAMP approach demonstrated significantly increased sensitivity through primer optimization and multiplexing.
- The developed RT-LAMP assays provide fast and reliable identification of SARS-CoV-2 infected individuals.
Conclusions:
- One-step multiplexed RT-LAMP assays are a highly promising option for developing easy-to-use and cost-effective POC test kits.
- This approach addresses the limitations of standard PCR for rapid, widespread SARS-CoV-2 screening.
