TY - JOUR
T1 - Homebrew
T2 - an economical and sensitive glassmilk-based nucleic-acid extraction method for SARS-CoV-2 diagnostics
AU - Page, Robert
AU - Scourfield, Edward
AU - Ficarelli, Mattia
AU - McKellar, Stuart
AU - Lee, Kwok Leung
AU - Maguire, Thomas J. A.
AU - Bouton, Clement
AU - Lista, Maria Jose
AU - Neil, Stuart J. D.
AU - Malim, Michael H.
AU - Zuckerman, Mark
AU - Mischo, Hannah E.
AU - Martinez-Nunez, Rocio
N1 - Funding Information:
This work was funded by a King’s Together Rapid COVID-19 Call award to R.T.M.-N. and S.J.D.N. and a Huo Family Foundation Award to M.H.M., R.T.M.-N., and S.J.D.N. H.E.M. and S.M. were funded by a Sir Henry Dale Fellowship jointly funded by the Wellcome Trust and the Royal Society ( 218537/Z/19/Z ) to H.E.M. R.T.M.-N. was supported by the Wellcome Trust ( 213984/Z/18/Z ). R.P. was supported through the ( NIHR ) Biomedical Research Centre based at Guy’s and St. Thomas’ NHS Foundation Trust in partnership with King’s College London and King’s College Hospital NHS Foundation Trust . M.F. was supported by the MRC-KCL Doctoral Training Partnership in Biomedical Sciences ( MR/N013700/1 ) and MRC ( MR/R50225X/1 ). This research was funded in whole, or in part, by the Wellcome Trust 218537/Z/19/Z and 213984/Z/18/Z. For the purpose of open access, the author has applied a CC BY public copyright license to any author accepted manuscript version arising from this submission.
Funding Information:
This work was funded by a King's Together Rapid COVID-19 Call award to R.T.M.-N. and S.J.D.N. and a Huo Family Foundation Award to M.H.M. R.T.M.-N. and S.J.D.N. H.E.M. and S.M. were funded by a Sir Henry Dale Fellowship jointly funded by the Wellcome Trust and the Royal Society (218537/Z/19/Z) to H.E.M. R.T.M.-N. was supported by the Wellcome Trust (213984/Z/18/Z). R.P. was supported through the (NIHR) Biomedical Research Centre based at Guy's and St. Thomas? NHS Foundation Trust in partnership with King's College London and King's College Hospital NHS Foundation Trust. M.F. was supported by the MRC-KCL Doctoral Training Partnership in Biomedical Sciences (MR/N013700/1) and MRC (MR/R50225X/1). This research was funded in whole, or in part, by the Wellcome Trust 218537/Z/19/Z and 213984/Z/18/Z. For the purpose of open access, the author has applied a CC BY public copyright license to any author accepted manuscript version arising from this submission. R.P. E.S. M.F. S.M. K.L.L. T.J.A.M. C.B. and M.J.L. performed experiments. S.J.D.N. M.H.M. and M.Z. provided intellectual input and samples. H.E.M. and R.T.M.-N. supervised the project and designed and performed experiments. R.P. H.E.M. and R.T.M.-N. wrote the initial manuscript; all authors were involved in editing the manuscript. The authors declare no competing interests. The author list of this paper includes contributors from the location where the research was conducted who participated in the data collection, design, analysis, and/or interpretation of the work.
Publisher Copyright:
© 2022 The Authors
PY - 2022/3/28
Y1 - 2022/3/28
N2 - Management of COVID-19 and other epidemics requires large-scale diagnostic testing. The gold standard for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection remains reverse transcription quantitative PCR (qRT-PCR) analysis, which detects viral RNA more sensitively than any other method. However, the resource use and supply-chain requirements of RT-PCR have continued to challenge diagnostic laboratories worldwide. Here, we establish and characterize a low-cost method to detect SARS-CoV-2 in clinical combined nose and throat swabs, allowing for automation in high-throughput settings. This method inactivates virus material with sodium dodecylsulfate (SDS) and uses silicon dioxide as the RNA-binding matrix in combination with sodium chloride (NaCl) and isopropanol. With similar sensitivity for SARS-CoV-2 viral targets but a fraction of time and reagent expenditure compared with commercial kits, our method also enables sample pooling without loss of sensitivity. We suggest that this method will facilitate more economical widespread testing, particularly in resource-limited settings.
AB - Management of COVID-19 and other epidemics requires large-scale diagnostic testing. The gold standard for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection remains reverse transcription quantitative PCR (qRT-PCR) analysis, which detects viral RNA more sensitively than any other method. However, the resource use and supply-chain requirements of RT-PCR have continued to challenge diagnostic laboratories worldwide. Here, we establish and characterize a low-cost method to detect SARS-CoV-2 in clinical combined nose and throat swabs, allowing for automation in high-throughput settings. This method inactivates virus material with sodium dodecylsulfate (SDS) and uses silicon dioxide as the RNA-binding matrix in combination with sodium chloride (NaCl) and isopropanol. With similar sensitivity for SARS-CoV-2 viral targets but a fraction of time and reagent expenditure compared with commercial kits, our method also enables sample pooling without loss of sensitivity. We suggest that this method will facilitate more economical widespread testing, particularly in resource-limited settings.
UR - http://www.scopus.com/inward/record.url?scp=85126921315&partnerID=8YFLogxK
U2 - 10.1016/j.crmeth.2022.100186
DO - 10.1016/j.crmeth.2022.100186
M3 - Article
VL - 2
JO - Cell Reports Methods
JF - Cell Reports Methods
IS - 3
M1 - 100186
ER -