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

The Hebrew University-Hadassah COVID-19 Diagnosis Team

Research output: Contribution to journalArticlepeer-review

141 Scopus citations


Introduction: Testing for active SARS-CoV-2 infection is a fundamental tool in the public health measures taken to control the COVID-19 pandemic. Because of the overwhelming use of SARS-CoV-2 reverse transcription (RT)-PCR tests worldwide, the availability of test kits has become a major bottleneck and the need to increase testing throughput is rising. We aim to overcome these challenges by pooling samples together, and performing RNA extraction and RT-PCR in pools. Methods: We tested the efficiency and sensitivity of pooling strategies for RNA extraction and RT-PCR detection of SARS-CoV-2. We tested 184 samples both individually and in pools to estimate the effects of pooling. We further implemented Dorfman pooling with a pool size of eight samples in large-scale clinical tests. Results: We demonstrated pooling strategies that increase testing throughput while maintaining high sensitivity. A comparison of 184 samples tested individually and in pools of eight samples showed that test results were not significantly affected. Implementing the eight-sample Dorfman pooling to test 26 576 samples from asymptomatic individuals, we identified 31 (0.12%) SARS-CoV-2 positive samples, achieving a 7.3-fold increase in throughput. Discussion: Pooling approaches for SARS-CoV-2 testing allow a drastic increase in throughput while maintaining clinical sensitivity. We report the successful large-scale pooled screening of asymptomatic populations.

Original languageEnglish
Pages (from-to)1248-1253
Number of pages6
JournalClinical Microbiology and Infection
Issue number9
StatePublished - Sep 2020

Bibliographical note

Publisher Copyright:
© 2020 European Society of Clinical Microbiology and Infectious Diseases


  • COVID-19
  • Diagnostics
  • Group testing
  • Infectious diseases
  • RT-PCR


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