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Improved sensitivity, safety, and rapidity of COVID-19 tests by replacing viral storage solution with lysis buffer

Conducting numerous, rapid, and reliable PCR tests for SARS-CoV-2 is essential for our ability to monitor and control the current COVID-19 pandemic. Here, we tested the sensitivity and efficiency of SARS-CoV-2 detection in clinical samples collected directly into a mix of lysis buffer and RNA preser...

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Autores principales: Erster, Oran, Shkedi, Omer, Benedek, Gil, Zilber, Eyal, Varkovitzky, Itay, Shirazi, Rachel, Oriya Shorka, Dorit, Cohen, Yuval, Bar, Tzahi, Yechieli, Rafi, Tepperberg Oikawa, Michal, Venkert, Dana, Linial, Michal, Oiknine-Djian, Esther, Mandelboim, Michal, Livneh, Zvi, Shenhav-Saltzman, Gilat, Mendelson, Ella, Wolf, Dana, Szwarcwort-Cohen, Moran, Mor, Orna, Lewis, Yair, Zeevi, Danny
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8009371/
https://www.ncbi.nlm.nih.gov/pubmed/33784369
http://dx.doi.org/10.1371/journal.pone.0249149
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author Erster, Oran
Shkedi, Omer
Benedek, Gil
Zilber, Eyal
Varkovitzky, Itay
Shirazi, Rachel
Oriya Shorka, Dorit
Cohen, Yuval
Bar, Tzahi
Yechieli, Rafi
Tepperberg Oikawa, Michal
Venkert, Dana
Linial, Michal
Oiknine-Djian, Esther
Mandelboim, Michal
Livneh, Zvi
Shenhav-Saltzman, Gilat
Mendelson, Ella
Wolf, Dana
Szwarcwort-Cohen, Moran
Mor, Orna
Lewis, Yair
Zeevi, Danny
author_facet Erster, Oran
Shkedi, Omer
Benedek, Gil
Zilber, Eyal
Varkovitzky, Itay
Shirazi, Rachel
Oriya Shorka, Dorit
Cohen, Yuval
Bar, Tzahi
Yechieli, Rafi
Tepperberg Oikawa, Michal
Venkert, Dana
Linial, Michal
Oiknine-Djian, Esther
Mandelboim, Michal
Livneh, Zvi
Shenhav-Saltzman, Gilat
Mendelson, Ella
Wolf, Dana
Szwarcwort-Cohen, Moran
Mor, Orna
Lewis, Yair
Zeevi, Danny
author_sort Erster, Oran
collection PubMed
description Conducting numerous, rapid, and reliable PCR tests for SARS-CoV-2 is essential for our ability to monitor and control the current COVID-19 pandemic. Here, we tested the sensitivity and efficiency of SARS-CoV-2 detection in clinical samples collected directly into a mix of lysis buffer and RNA preservative, thus inactivating the virus immediately after sampling. We tested 79 COVID-19 patients and 20 healthy controls. We collected two samples (nasopharyngeal swabs) from each participant: one swab was inserted into a test tube with Viral Transport Medium (VTM), following the standard guideline used as the recommended method for sample collection; the other swab was inserted into a lysis buffer supplemented with nucleic acid stabilization mix (coined NSLB). We found that RT-qPCR tests of patients were significantly more sensitive with NSLB sampling, reaching detection threshold 2.1±0.6 (Mean±SE) PCR cycles earlier then VTM samples from the same patient. We show that this improvement is most likely since NSLB samples are not diluted in lysis buffer before RNA extraction. Re-extracting RNA from NSLB samples after 72 hours at room temperature did not affect the sensitivity of detection, demonstrating that NSLB allows for long periods of sample preservation without special cooling equipment. We also show that swirling the swab in NSLB and discarding it did not reduce sensitivity compared to retaining the swab in the tube, thus allowing improved automation of COVID-19 tests. Overall, we show that using NSLB instead of VTM can improve the sensitivity, safety, and rapidity of COVID-19 tests at a time most needed.
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spelling pubmed-80093712021-04-07 Improved sensitivity, safety, and rapidity of COVID-19 tests by replacing viral storage solution with lysis buffer Erster, Oran Shkedi, Omer Benedek, Gil Zilber, Eyal Varkovitzky, Itay Shirazi, Rachel Oriya Shorka, Dorit Cohen, Yuval Bar, Tzahi Yechieli, Rafi Tepperberg Oikawa, Michal Venkert, Dana Linial, Michal Oiknine-Djian, Esther Mandelboim, Michal Livneh, Zvi Shenhav-Saltzman, Gilat Mendelson, Ella Wolf, Dana Szwarcwort-Cohen, Moran Mor, Orna Lewis, Yair Zeevi, Danny PLoS One Research Article Conducting numerous, rapid, and reliable PCR tests for SARS-CoV-2 is essential for our ability to monitor and control the current COVID-19 pandemic. Here, we tested the sensitivity and efficiency of SARS-CoV-2 detection in clinical samples collected directly into a mix of lysis buffer and RNA preservative, thus inactivating the virus immediately after sampling. We tested 79 COVID-19 patients and 20 healthy controls. We collected two samples (nasopharyngeal swabs) from each participant: one swab was inserted into a test tube with Viral Transport Medium (VTM), following the standard guideline used as the recommended method for sample collection; the other swab was inserted into a lysis buffer supplemented with nucleic acid stabilization mix (coined NSLB). We found that RT-qPCR tests of patients were significantly more sensitive with NSLB sampling, reaching detection threshold 2.1±0.6 (Mean±SE) PCR cycles earlier then VTM samples from the same patient. We show that this improvement is most likely since NSLB samples are not diluted in lysis buffer before RNA extraction. Re-extracting RNA from NSLB samples after 72 hours at room temperature did not affect the sensitivity of detection, demonstrating that NSLB allows for long periods of sample preservation without special cooling equipment. We also show that swirling the swab in NSLB and discarding it did not reduce sensitivity compared to retaining the swab in the tube, thus allowing improved automation of COVID-19 tests. Overall, we show that using NSLB instead of VTM can improve the sensitivity, safety, and rapidity of COVID-19 tests at a time most needed. Public Library of Science 2021-03-30 /pmc/articles/PMC8009371/ /pubmed/33784369 http://dx.doi.org/10.1371/journal.pone.0249149 Text en © 2021 Erster et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Erster, Oran
Shkedi, Omer
Benedek, Gil
Zilber, Eyal
Varkovitzky, Itay
Shirazi, Rachel
Oriya Shorka, Dorit
Cohen, Yuval
Bar, Tzahi
Yechieli, Rafi
Tepperberg Oikawa, Michal
Venkert, Dana
Linial, Michal
Oiknine-Djian, Esther
Mandelboim, Michal
Livneh, Zvi
Shenhav-Saltzman, Gilat
Mendelson, Ella
Wolf, Dana
Szwarcwort-Cohen, Moran
Mor, Orna
Lewis, Yair
Zeevi, Danny
Improved sensitivity, safety, and rapidity of COVID-19 tests by replacing viral storage solution with lysis buffer
title Improved sensitivity, safety, and rapidity of COVID-19 tests by replacing viral storage solution with lysis buffer
title_full Improved sensitivity, safety, and rapidity of COVID-19 tests by replacing viral storage solution with lysis buffer
title_fullStr Improved sensitivity, safety, and rapidity of COVID-19 tests by replacing viral storage solution with lysis buffer
title_full_unstemmed Improved sensitivity, safety, and rapidity of COVID-19 tests by replacing viral storage solution with lysis buffer
title_short Improved sensitivity, safety, and rapidity of COVID-19 tests by replacing viral storage solution with lysis buffer
title_sort improved sensitivity, safety, and rapidity of covid-19 tests by replacing viral storage solution with lysis buffer
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8009371/
https://www.ncbi.nlm.nih.gov/pubmed/33784369
http://dx.doi.org/10.1371/journal.pone.0249149
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