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One-tube SARS-CoV-2 detection platform based on RT-RPA and CRISPR/Cas12a
Yangyang Sun1,2,3,4, Lei Yu1,2,3,4, Chengxi Liu5
1Department of Urology, International Cancer Center, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, Shenzhen University School of Medicine, Shenzhen, 518039, China.
Journal of Translational Medicine
|February 17, 2021
Summary
A new OR-DETECTR test offers rapid and accurate detection of SARS-CoV-2, the virus causing COVID-19. This one-tube molecular assay is suitable for widespread surveillance and diagnosis.
Area of Science:
- Molecular Biology
- Infectious Disease Diagnostics
- Biotechnology
Background:
- The COVID-19 pandemic necessitates robust surveillance systems for SARS-CoV-2 detection.
- Nucleic acid detection is crucial for effective SARS-CoV-2 monitoring, especially when easing public health measures.
Purpose of the Study:
- To develop a rapid, one-tube detection platform for SARS-CoV-2 using RT-RPA and CRISPR-DETECTR technology.
- To create an accessible and efficient diagnostic tool for COVID-19 detection.
Main Methods:
- Developed the OR-DETECTR platform combining Reverse Transcription and Recombinase Polymerase Isothermal Amplification (RT-RPA) with DNA Endonuclease-Targeted CRISPR Trans Reporter (DETECTR).
- Designed specific primers for SARS-CoV-2 RdRp and N genes and optimized a one-tube reaction.
- Validated specificity against other human coronaviruses and Influenza A, and assessed clinical samples against rRT-PCR.
Main Results:
- The OR-DETECTR platform provides results in approximately 50 minutes with high specificity and a low detection limit (2.5 copies/µl RNA standard).
- Clinical validation showed 100% consistency with rRT-PCR, accurately identifying SARS-CoV-2 positive and negative samples.
- A lateral flow assay version of OR-DETECTR was developed for simplified COVID-19 detection.
Conclusions:
- OR-DETECTR is a rapid, accurate, and user-friendly molecular diagnostic platform for SARS-CoV-2.
- The system's ease of use and lack of requirement for large instruments make it suitable for diverse testing settings.
- The developed platform, including a lateral flow assay, supports effective COVID-19 surveillance and diagnosis.
