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Cas12a-assisted RTF-EXPAR for accurate, rapid and simple detection of SARS-CoV-2 RNA
Xiao-Min Hang1, Hui-Yi Wang1, Peng-Fei Liu2
1School of Pharmacy, Jiangsu University, Zhenjiang, 212013, PR China.
Biosensors & Bioelectronics
|September 11, 2022
Summary
A new Cas12a-assisted RTF-EXPAR strategy rapidly identifies SARS-CoV-2 RNA within 40 minutes. This ultrasensitive method offers accurate, point-of-care detection with minimal equipment, aiding COVID-19 pandemic control.
Area of Science:
- Molecular Biology
- Biotechnology
- Infectious Disease Diagnostics
Background:
- Rapid and accurate identification of SARS-CoV-2 is crucial for controlling the COVID-19 pandemic.
- Existing diagnostic methods may require complex equipment or lengthy procedures.
Purpose of the Study:
- To develop and evaluate a fast, sensitive, and accurate method for SARS-CoV-2 RNA detection.
- To assess the performance of a Cas12a-assisted RTF-EXPAR strategy for point-of-care applications.
Main Methods:
- Combined Reverse Transcriptase-Fuelled Exponential Amplification Reaction (RTF-EXPAR) with CRISPR-Cas12a.
- Utilized isothermal control for amplification and detection.
- Employed end-point fluorescence and smartphone-assisted RGB analysis for readout.
Main Results:
- Detected SARS-CoV-2 RNA within 40 minutes.
- Achieved ultrasensitive detection limits of 3.77 aM (∼2 copies/μL) via fluorescence and 4.81 aM (∼3 copies/μL) via smartphone analysis.
- Cas12a enhanced specificity through intrinsic sequence-specific recognition.
Conclusions:
- The Cas12a-assisted RTF-EXPAR strategy is a fast, sensitive, and accurate method for SARS-CoV-2 RNA detection.
- The assay requires minimal equipment, making it suitable for point-of-care molecular diagnostics.
- This approach shows significant promise for rapid screening and management of COVID-19.
