Rapid isothermal amplification and portable detection system for SARS-CoV-2

Area of Science:

• Molecular Biology
• Virology
• Biotechnology

Background:

• The COVID-19 pandemic highlighted critical needs for accessible diagnostics.
• Conventional laboratory infrastructure limits rapid SARS-CoV-2 detection.
• Established methods like RT-PCR require specialized equipment and trained personnel.

Purpose of the Study:

• To develop and validate a rapid, point-of-care (POC) isothermal assay for SARS-CoV-2 detection.
• To demonstrate the assay's performance on clinical samples using a portable instrument.
• To provide an alternative diagnostic pathway for resource-limited settings.

Main Methods:

• Developed a reverse transcription loop mediated isothermal amplification (RT-LAMP) assay for SARS-CoV-2.
• Characterized the assay's limit of detection (LOD) using spiked synthetic nasal fluid.
• Validated the assay on clinical samples using a portable POC device with a smartphone reader.
• Performed RT-LAMP directly on viral transport medium (VTM) without RNA extraction.

Main Results:

• The RT-LAMP assay achieved an LOD of 50 RNA copies/μL in VTM within 30 minutes.
• The assay successfully detected SARS-CoV-2 in 20 clinical samples.
• The portable POC system accurately identified SARS-CoV-2 in 10 clinical samples, agreeing with RT-PCR controls.
• The POC device provided results within 30 minutes.

Conclusions:

• An accessible RT-LAMP assay and POC system can reliably detect SARS-CoV-2.
• This diagnostic approach bypasses the need for conventional laboratory infrastructure.
• The developed method offers a viable solution for point-of-sample-collection diagnostics.