CRISPR-Cas12-based detection of SARS-CoV-2

Area of Science:

• Molecular Biology
• Infectious Diseases
• Biotechnology

Background:

• The COVID-19 pandemic, caused by SARS-CoV-2, has necessitated rapid and accurate diagnostic tools.
• Existing methods like RT-PCR, while accurate, can be time-consuming and require specialized equipment.
• There is a need for faster, easily implementable diagnostic assays for SARS-CoV-2 detection.

Purpose of the Study:

• To develop and validate a rapid, easy-to-implement CRISPR-Cas12-based lateral flow assay for SARS-CoV-2 detection.
• To compare the performance of the CRISPR-based assay against established RT-PCR methods.

Main Methods:

• Development of a CRISPR-Cas12-based lateral flow assay (DETECTR) for detecting SARS-CoV-2 RNA from respiratory swab extracts.
• Validation using contrived reference samples and clinical samples from patients in the United States.
• Comparison with the CDC's real-time RT-PCR assay.

Main Results:

• The CRISPR-based DETECTR assay provides results in under 40 minutes.
• The assay demonstrated 95% positive predictive agreement and 100% negative predictive agreement when compared to RT-PCR.
• The assay was validated using samples from 36 COVID-19 patients and 42 patients with other respiratory infections.

Conclusions:

• The CRISPR-Cas12-based DETECTR assay is a rapid, accurate, and visually interpretable alternative for SARS-CoV-2 detection.
• This assay offers a significant improvement in speed and ease of implementation compared to traditional RT-PCR methods.
• The assay shows high predictive agreement, making it a viable tool for diagnosing COVID-19 infections.