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Massive and rapid COVID-19 testing is feasible by extraction-free SARS-CoV-2 RT-PCR.
Nat Commun. 2020 09 23; 11(1):4812.NC

Abstract

Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is commonly diagnosed by reverse transcription polymerase chain reaction (RT-PCR) to detect viral RNA in patient samples, but RNA extraction constitutes a major bottleneck in current testing. Methodological simplification could increase diagnostic availability and efficiency, benefitting patient care and infection control. Here, we describe methods circumventing RNA extraction in COVID-19 testing by performing RT-PCR directly on heat-inactivated or lysed samples. Our data, including benchmarking using 597 clinical patient samples and a standardised diagnostic system, demonstrate that direct RT-PCR is viable option to extraction-based tests. Using controlled amounts of active SARS-CoV-2, we confirm effectiveness of heat inactivation by plaque assay and evaluate various generic buffers as transport medium for direct RT-PCR. Significant savings in time and cost are achieved through RNA-extraction-free protocols that are directly compatible with established PCR-based testing pipelines. This could aid expansion of COVID-19 testing.

Authors+Show Affiliations

Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 171 77, Stockholm, Sweden.Department of Clinical Microbiology, Karolinska University Hospital, 171 76, Stockholm, Sweden.Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 171 77, Stockholm, Sweden.Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, 171 77, Stockholm, Sweden.Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 171 77, Stockholm, Sweden.Department of Clinical Microbiology, Karolinska University Hospital, 171 76, Stockholm, Sweden.Department of Clinical Microbiology, Karolinska University Hospital, 171 76, Stockholm, Sweden.Department of Clinical Microbiology, Karolinska University Hospital, 171 76, Stockholm, Sweden.Public Health Agency of Sweden, 171 82, Solna, Sweden.Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, 171 77, Stockholm, Sweden.Department of Clinical Microbiology, Karolinska University Hospital, 171 76, Stockholm, Sweden. Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, 171 77, Stockholm, Sweden.Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 171 77, Stockholm, Sweden.Department of Medical Biochemistry and Biophysics, Karolinska Institutet, 171 77, Stockholm, Sweden. bjorn.reinius@ki.se.

Pub Type(s)

Comparative Study
Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't
Validation Study

Language

eng

PubMed ID

32968075

Citation

Smyrlaki, Ioanna, et al. "Massive and Rapid COVID-19 Testing Is Feasible By Extraction-free SARS-CoV-2 RT-PCR." Nature Communications, vol. 11, no. 1, 2020, p. 4812.
Smyrlaki I, Ekman M, Lentini A, et al. Massive and rapid COVID-19 testing is feasible by extraction-free SARS-CoV-2 RT-PCR. Nat Commun. 2020;11(1):4812.
Smyrlaki, I., Ekman, M., Lentini, A., Rufino de Sousa, N., Papanicolaou, N., Vondracek, M., Aarum, J., Safari, H., Muradrasoli, S., Rothfuchs, A. G., Albert, J., Högberg, B., & Reinius, B. (2020). Massive and rapid COVID-19 testing is feasible by extraction-free SARS-CoV-2 RT-PCR. Nature Communications, 11(1), 4812. https://doi.org/10.1038/s41467-020-18611-5
Smyrlaki I, et al. Massive and Rapid COVID-19 Testing Is Feasible By Extraction-free SARS-CoV-2 RT-PCR. Nat Commun. 2020 09 23;11(1):4812. PubMed PMID: 32968075.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Massive and rapid COVID-19 testing is feasible by extraction-free SARS-CoV-2 RT-PCR. AU - Smyrlaki,Ioanna, AU - Ekman,Martin, AU - Lentini,Antonio, AU - Rufino de Sousa,Nuno, AU - Papanicolaou,Natali, AU - Vondracek,Martin, AU - Aarum,Johan, AU - Safari,Hamzah, AU - Muradrasoli,Shaman, AU - Rothfuchs,Antonio Gigliotti, AU - Albert,Jan, AU - Högberg,Björn, AU - Reinius,Björn, Y1 - 2020/09/23/ PY - 2020/05/12/received PY - 2020/09/02/accepted PY - 2020/9/24/entrez PY - 2020/9/25/pubmed PY - 2020/10/2/medline SP - 4812 EP - 4812 JF - Nature communications JO - Nat Commun VL - 11 IS - 1 N2 - Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is commonly diagnosed by reverse transcription polymerase chain reaction (RT-PCR) to detect viral RNA in patient samples, but RNA extraction constitutes a major bottleneck in current testing. Methodological simplification could increase diagnostic availability and efficiency, benefitting patient care and infection control. Here, we describe methods circumventing RNA extraction in COVID-19 testing by performing RT-PCR directly on heat-inactivated or lysed samples. Our data, including benchmarking using 597 clinical patient samples and a standardised diagnostic system, demonstrate that direct RT-PCR is viable option to extraction-based tests. Using controlled amounts of active SARS-CoV-2, we confirm effectiveness of heat inactivation by plaque assay and evaluate various generic buffers as transport medium for direct RT-PCR. Significant savings in time and cost are achieved through RNA-extraction-free protocols that are directly compatible with established PCR-based testing pipelines. This could aid expansion of COVID-19 testing. SN - 2041-1723 UR - https://www.unboundmedicine.com/medline/citation/32968075/Massive_and_rapid_COVID_19_testing_is_feasible_by_extraction_free_SARS_CoV_2_RT_PCR_ L2 - https://doi.org/10.1038/s41467-020-18611-5 DB - PRIME DP - Unbound Medicine ER -