Unknown

Dataset Information

0

Massive and rapid COVID-19 testing is feasible by extraction-free SARS-CoV-2 RT-PCR.


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.

SUBMITTER: Smyrlaki I 

PROVIDER: S-EPMC7511968 | biostudies-literature | 2020 Sep

REPOSITORIES: biostudies-literature

altmetric image

Publications


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  ...[more]

Similar Datasets

| S-EPMC7927279 | biostudies-literature
| S-EPMC8175123 | biostudies-literature
| S-EPMC7472728 | biostudies-literature
| S-EPMC7405857 | biostudies-literature
| S-EPMC8408542 | biostudies-literature
| S-EPMC7885624 | biostudies-literature
| S-EPMC7598596 | biostudies-literature
| S-EPMC8349479 | biostudies-literature
| S-EPMC8230941 | biostudies-literature
| S-EPMC7924294 | biostudies-literature