Bitrus Inuwa

An international, interlaboratory ring trial confirms the feasibility of an extraction-less “direct” rt-qpcr method for reliable detection of sars-cov-2 rna in clinical samples

• Authors Details :  
• Margaret G. Mills,  
• Emily Bruce,  
• Meei-li Huang,  
• Jessica W,  
• Crothers,  
• Ollivier Hyrien,  
• Christopher A. L. Oura,  
• Lemar Blake,  
• Arianne Brown Jordan,  
• Susan Hester,  
• Leah Wehmas,  
• Bernard Mari,  
• Pascal Barby,  
• Caroline Lacoux,  
• Julien Fassy

Journal title : PLOS ONE

Publisher : Public Library of Science (PLoS)

Online ISSN : 1932-6203

Page Number : e0261853

Journal volume : 17

Journal issue : 1

216 Views Original Article

Reverse transcription–quantitative polymerase chain reaction (RT-qPCR) is used worldwide to test and trace the spread of severe acute respiratory syndrome coronavirus 2 (SARSCoV-2). “Extraction-less” or “direct” real time–reverse transcription polymerase chain reaction (RT-PCR) is a transparent and accessible qualitative method for SARS-CoV-2 detection from nasopharyngeal or oral pharyngeal samples with the potential to generate actionable data more quickly, at a lower cost, and with fewer experimental resources than full RT-qPCR. This study engaged 10 global testing sites, including laboratories currently experiencing testing limitations due to reagent or equipment shortages, in an international interlaboratory ring trial. Participating laboratories were provided a common protocol, common reagents, aliquots of identical pooled clinical samples, and purified nucleic acids and used their existing in-house equipment. We observed 100% concordance across laboratories in the correct identification of all positive and negative samples, with highly similar cycle threshold values. The test also performed well when applied to locally collected patient nasopharyngeal samples, provided the viral transport media did not contain charcoal or guanidine, both of which appeared to potently inhibit the RT-PCR reaction. Our results suggest that direct RT-PCR assay methods can be clearly translated across sites utilizing readily available equipment and expertise and are thus a feasible option for more efficient COVID-19 coronavirus disease testing as demanded by the continuing pandemic.

Article DOI & Crossmark Data

DOI : https://doi.org/10.1371/journal.pone.0261853

Article Subject Details

• Coronavirus
• Coronavirus disease
• Pandemic
• Public health

Article Keywords Details

• Coronavirus
• Direct RT-qPCR
• Extraction-less
• Interlaboratory ring trial
• Pandemic

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