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Massive and rapid C...
Massive and rapid COVID-19 testing is feasible by extraction-free SARS-CoV-2 RT-PCR
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- Smyrlaki, I (author)
- Karolinska Institutet
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Ekman, M (author)
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- Lentini, A (author)
- Karolinska Institutet
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- de Sousa, NR (author)
- Karolinska Institutet
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- Papanicolaou, N (author)
- Karolinska Institutet
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Vondracek, M (author)
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Aarum, J (author)
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Safari, H (author)
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Muradrasoli, S (author)
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- Rothfuchs, AG (author)
- Karolinska Institutet
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- Albert, J (author)
- Karolinska Institutet
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- Hogberg, B (author)
- Karolinska Institutet
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- Reinius, B (author)
- Karolinska Institutet
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(creator_code:org_t)
- 2020-09-23
- 2020
- English.
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In: Nature communications. - : Springer Science and Business Media LLC. - 2041-1723. ; 11:1, s. 4812-
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Abstract
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- 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.
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- ref (subject category)
- art (subject category)
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- By the author/editor
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Smyrlaki, I
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Ekman, M
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Lentini, A
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de Sousa, NR
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Papanicolaou, N
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Vondracek, M
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show more...
-
Aarum, J
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Safari, H
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Muradrasoli, S
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Rothfuchs, AG
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Albert, J
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Hogberg, B
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Reinius, B
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show less...
- Articles in the publication
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Nature communica ...
- By the university
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Karolinska Institutet