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

2020; Elsevier BV; Volume: 26; Issue: 9 Linguagem: Inglês

10.1016/j.cmi.2020.06.009

1469-0691

Roni Ben‐Ami, Agnes Klochendler, Michael Seidel, Tal Sido, Ori Gurel-Gurevich, Moran Yassour, Eran Meshorer, Gil Benedek, Irit Fogel, Esther Oiknine‐Djian, Arieh Gertler, Zeev Rotstein, Bruno Lavi, Yuval Dor, Dana G. Wolf, Maayan Salton, Yotam Drier, Agnes Klochendler, Amir Eden, Amar J. S. Klar, A. Geldman, Ayelet Arbel, Ayelet Peretz, Batel Shalom, Bracha Lea Ochana, D. Avrahami-Tzfati, Daniel Neiman, David Steinberg, D. Ben Zvi, Etai Shpigel, Gal Atlan, Harvey G. Klein, H. Chekroun, Hagit Shani, Itay Hazan, Irfan Ahmad Ansari, I. Magenheim, J. Moss, Judith Magenheim, Lior Peretz, L. Feigin, M. Saraby, Melanie A. Sherman, Michelle Bentata, Michel Avital, Marylee M. Kott, M. Reuben Peyser, Martin Weitz, Mordechai Shacham, Maria Grünewald, Noah J. Sasson, N. Wallis, Narmen Azazmeh, N. Tzarum, O. Fridlich, R Sher, Reba Condiotti, Ron Refaeli, Ronen Ben‐Ami, R. Zaken-Gallili, R. Gayman Helman, S. Ofek, S. Tzaban, Sheina Piyanzin, S. Anzi, Shlomo Dagan, S. Lilenthal, Tal Sido, Tamar Licht, T. Friehmann, Yotam Kaufman, Alexandre R.R. Péry, Ann Saada, Arie Dekel, A. Yeffet, Avraham Shaag, Ayelet Michael-Gayego, Elizabeth Shay, E. Arbib, Hadil Onallah, Kerem Ben‐Meir, L. Levinzon, Leonor Cohen‐Daniel, L. Natan, Muhammad Hamdan, Mila Rivkin, M. Shwieki, Olesya Vorontsov, Rima Barsuk, R. Abramovitch, R. Gutorov, Salim Sirhan, Sanofar Abdeen, Y. Yachnin, Yutti Daitch,

Biosensors and Analytical Detection

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.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.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.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.