A sensitive and affordable multiplex RT-qPCR assay for SARS-CoV-2 detection

Area of Science:

• Molecular Biology
• Virology
• Infectious Diseases

Background:

• The COVID-19 pandemic necessitates sensitive, specific, and affordable diagnostic tests for SARS-CoV-2.
• Current RT-qPCR assays are sensitive but often expensive and face supply chain limitations.
• There is a need for cost-effective diagnostic solutions, especially for low-resource settings.

Purpose of the Study:

• To develop and validate a multiplex RT-qPCR assay for SARS-CoV-2 detection.
• To assess the assay's performance against commercial standards.
• To evaluate the assay's cost-effectiveness and potential for high-volume testing.

Main Methods:

• Development of a multiplex assay targeting SARS-CoV-2, including human (RPP30) and viral (PhHV-1) controls.
• Validation of the assay's performance compared to existing commercial kits.
• Analysis of sample variability and correlation between human and viral nucleic acid levels.

Main Results:

• The developed multiplex assay demonstrated performance comparable to commercial RT-qPCR kits.
• The assay achieved comparable results at a significantly reduced cost (approximately 10% of commercial assays).
• A statistically significant correlation was found between human nucleic acid levels and SARS-CoV-2 detection, highlighting the utility of the human control.

Conclusions:

• A robust and cost-effective multiplex RT-qPCR assay for SARS-CoV-2 has been established.
• The assay's affordability and inclusion of controls can enhance diagnostic capacity, particularly in resource-limited regions.
• The human control provides a quantitative measure of sample quality, potentially reducing false-negative rates.