UnCovid: A versatile, low-cost, and open-source protocol for SARS-CoV-2 RNA detection

Area of Science:

• Molecular Biology
• Virology
• Biotechnology

Background:

• The COVID-19 pandemic necessitates accessible and reliable diagnostic tools.
• Existing SARS-CoV-2 detection methods may require specialized equipment or complex procedures.
• There is a need for field-deployable assays for widespread surveillance.

Purpose of the Study:

• To develop and detail a robust protocol for SARS-CoV-2 RNA detection.
• To enable qualitative and quantitative detection using accessible molecular biology equipment.
• To adapt the protocol for use with the portable BentoLab molecular workstation for remote applications.

Main Methods:

• Reverse transcription-polymerase chain reaction (RT-PCR) for amplifying target RNA.
• CRISPR/Cas system for visualizing amplified SARS-CoV-2 genetic material.
• Detection of two specific SARS-CoV-2 genomic regions and a human RNaseP control.
• Utilized conventional thermocyclers, transilluminators, and fluorescence plate readers.

Main Results:

• A detailed, step-by-step protocol for SARS-CoV-2 RNA detection was established.
• The assay demonstrated capability for both qualitative and quantitative measurements.
• The protocol was successfully adapted for use with the portable BentoLab workstation.

Conclusions:

• The described RT-PCR and CRISPR/Cas-based protocol offers a versatile method for SARS-CoV-2 RNA detection.
• The assay's adaptability to portable equipment facilitates its use in resource-limited or remote settings.
• This protocol contributes to accessible diagnostic strategies for infectious diseases.