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Preliminary support for a "dry swab, extraction free" protocol for SARS-CoV-2 testing via RT-qPCR.

Sanjay Srivatsan, Peter D. Han, Katrina van Raay, Caitlin R. Wolf, Denise J. McCulloch, Ashley E. Kim, Elisabeth Brandstetter, Beth Martin, Jase Gehring, Wei Chen, Seattle Flu Study Investigators, Sriram Kosuri, Eric Q. Konnick, Christina M. Lockwood, Rieder, Mark J, Deborah A. Nickerson, Helen Y. Chu, Jay Shendure, Lea M. Starita

2020eScholarship (California Digital Library)46 citations

Abstract

The urgent need for massively scaled clinical or surveillance testing for SARS-CoV-2 has necessitated a reconsideration of the methods by which respiratory samples are collected, transported, processed and tested. Conventional testing for SARS-CoV-2 involves collection of a clinical specimen with a nasopharyngeal swab, storage of the swab during transport in universal transport medium (UTM), extraction of RNA, and quantitative reverse transcription PCR (RT-qPCR). As testing has scaled across the world, supply chain challenges have emerged across this entire workflow. Here we sought to evaluate how eliminating the UTM storage and RNA extraction steps would impact the results of molecular testing. Using paired mid-turbinate swabs self-collected by 11 individuals with previously established SARS-CoV-2 positivity, we performed a comparison of conventional (swab → UTM → RNA extraction → RT-qPCR) vs. simplified (direct elution from dry swab → RT-qPCR) protocols. Our results suggest that dry swabs eluted directly into a simple buffered solution (TE) can support molecular detection of SARS-CoV-2 via endpoint RT-qPCR without substantially compromising sensitivity. Although further confirmation with a larger sample size and variation of other parameters is necessary, these results are encouraging for the possibility of a simplified workflow that could support massively scaled testing for COVID-19 control.

Topics & Concepts

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)RNA extractionCoronavirus disease 2019 (COVID-19)RNABiologyMedicinePathologyGeneticsGeneInfectious disease (medical specialty)DiseaseSARS-CoV-2 detection and testingBiosensors and Analytical DetectionAdvanced biosensing and bioanalysis techniques
Preliminary support for a "dry swab, extraction free" protocol for SARS-CoV-2 testing via RT-qPCR. | Litcius