Extraction-free COVID-19 (SARS-CoV-2) diagnosis by RT-PCR to increase capacity for national testing programmes during a pandemic

Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2) causes Coronavirus disease 2019 (COVID-19), a respiratory tract infection. The standard molecular diagnostic test is a multistep process involving viral RNA extraction and real-time quantitative reverse transcriptase PCR (qRT-PCR). Laborat...

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Bibliographic Details
Published inbioRxiv
Main Authors Grant, Paul R, Turner, Melanie A, Gee Yen Shin, Nastouli, Eleni, Levett, Lisa J
Format Paper
LanguageEnglish
Published Cold Spring Harbor Cold Spring Harbor Laboratory Press 09.04.2020
Subjects
Coronaviruses
COVID-19
DNA probes
Pandemics
Polymerase chain reaction
Respiratory tract diseases
Ribonucleic acid
RNA
RNA-directed DNA polymerase
Severe acute respiratory syndrome coronavirus 2
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Summary:Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2) causes Coronavirus disease 2019 (COVID-19), a respiratory tract infection. The standard molecular diagnostic test is a multistep process involving viral RNA extraction and real-time quantitative reverse transcriptase PCR (qRT-PCR). Laboratories across the globe face constraints on equipment and reagents during the COVID-19 pandemic. We have developed a simplified qRT-PCR assay that removes the need for an RNA extraction process and can be run on a real-time thermal cycler. The assay uses custom primers and probes, and maintains diagnostic sensitivity within 98.0% compared to the assay run on a high-throughput, random-access automated platform, the Panther Fusion (Hologic). This assay can be used to increase capacity for COVID-19 testing for national programmes worldwide. Competing Interest Statement The authors have declared no competing interest. Footnotes * Added Panther Fusion SARS-CoV-2 method
DOI:10.1101/2020.04.06.028316