Large-scale implementation of pooled RNA extraction and RT-PCR for SARS-CoV-2 detection.

Area of Science:

• Molecular Biology
• Virology
• Public Health

Background:

• Reverse transcription polymerase chain reaction (RT-PCR) is crucial for detecting SARS-CoV-2.
• High demand for RT-PCR tests has created bottlenecks in global testing capacity.
• Increased testing throughput is essential for managing the COVID-19 pandemic.

Purpose of the Study:

• To evaluate the efficiency and sensitivity of sample pooling for SARS-CoV-2 RNA extraction and RT-PCR.
• To address limitations in testing kit availability and increase overall testing throughput.

Main Methods:

• Tested 184 samples individually and in pools to assess pooling effects on RNA extraction and RT-PCR sensitivity.
• Implemented Dorfman pooling strategy with a pool size of eight samples for large-scale clinical testing.

Main Results:

• Pooling strategies were demonstrated to enhance testing throughput while maintaining high sensitivity.
• Testing 184 samples individually versus in pools of eight showed no significant impact on test results.
• Large-scale screening of 26,576 asymptomatic individuals using eight-sample pools identified 31 positive SARS-CoV-2 cases, achieving a 7.3-fold throughput increase.

Conclusions:

• Sample pooling for SARS-CoV-2 detection drastically increases testing throughput.
• Pooling maintains clinical sensitivity, enabling effective screening of asymptomatic populations.