Evaluation of COVID-19 RT-qPCR Test in Multi sample Pools

Area of Science:

• Virology
• Molecular Biology
• Public Health

Background:

• The COVID-19 pandemic, caused by SARS-CoV-2, has strained healthcare systems globally.
• Reagent shortages for diagnostic tests hinder effective pandemic response.
• There is an urgent need for scalable and efficient SARS-CoV-2 detection protocols.

Purpose of the Study:

• To evaluate the feasibility of a sample pooling strategy for SARS-CoV-2 detection.
• To determine the maximum pool size for reliable detection using RT-qPCR.
• To assess the applicability of pooling in clinical settings.

Main Methods:

• RNA extracts and transport media from positive samples were pooled with negative samples.
• Pools of varying sizes (2- to 64-samples) were tested using reverse transcription quantitative polymerase chain reaction (RT-qPCR).
• Pooling was performed both before and after RNA extraction.

Main Results:

• A single positive sample was reliably detected in pools of up to 32 samples.
• Detection in pools of up to 64 samples may be possible with adjusted amplification cycles.
• Pooling is effective regardless of whether it occurs before or after RNA extraction.

Conclusions:

• Sample pooling is an immediately applicable strategy for increasing SARS-CoV-2 screening capacity in clinical laboratories.
• This method utilizes standard reagents and equipment, facilitating rapid implementation.
• Expanding screening capacity through pooling can enhance community surveillance and targeted outbreak detection in high-risk groups.