Processing Hundreds of SARS-CoV-2 Samples with an In-House PCR-Based Method without Robotics

Area of Science:

• Molecular Biology
• Virology
• Clinical Diagnostics

Background:

• The COVID-19 pandemic necessitated rapid development of SARS-CoV-2 testing systems.
• Existing diagnostic protocols often require specialized equipment or complex procedures.

Purpose of the Study:

• To develop an efficient, non-robotic PCR test for screening COVID-19 infections.
• To optimize RNA extraction and RT-qPCR procedures for high throughput.
• To validate sample stability and compatibility with various testing platforms.

Main Methods:

• Developed a streamlined protocol using multichannel pipettes and magnetic bead-based RNA extraction.
• Combined reverse transcription and quantitative PCR (RT-qPCR) into a single step.
• Evaluated two RT-qPCR kits, various sampling buffers (NaCl, PBS), and a novel human RNase P internal control.

Main Results:

• The protocol can process up to ~180 samples per day without robotics.
• Samples collected in NaCl or PBS buffers are stable and compatible with COVID-19 testing.
• The developed internal control functions effectively without a DNA digestion step.
• The test achieves sensitivity and accuracy comparable to standardized clinical diagnostic protocols.

Conclusions:

• A simplified, high-throughput PCR test for SARS-CoV-2 detection has been established.
• The protocol is user-friendly and suitable for clinical settings.
• This method offers a viable alternative for large-scale COVID-19 screening.