Inactivation of SARS-CoV-2 virus in saliva using a guanidium based transport medium suitable for RT-PCR diagnostic assays.

<h4>Background</h4>Upper respiratory samples used to test for SARS-CoV-2 virus may be infectious and present a hazard during transport and testing. A buffer with the ability to inactivate SARS-CoV-2 at the time of sample collection could simplify and expand testing for COVID-19 to non-co...

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Autores principales:	Sukalyani Banik, Kaheerman Saibire, Shraddha Suryavanshi, Glenn Johns, Soumitesh Chakravorty, Robert Kwiatkowski, David Alland, Padmapriya P Banada
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/aac5e2b0b2c6472c9b2acee91bd089af
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spelling	oai:doaj.org-article:aac5e2b0b2c6472c9b2acee91bd089af2021-12-02T20:03:53ZInactivation of SARS-CoV-2 virus in saliva using a guanidium based transport medium suitable for RT-PCR diagnostic assays.1932-620310.1371/journal.pone.0252687https://doaj.org/article/aac5e2b0b2c6472c9b2acee91bd089af2021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0252687https://doaj.org/toc/1932-6203<h4>Background</h4>Upper respiratory samples used to test for SARS-CoV-2 virus may be infectious and present a hazard during transport and testing. A buffer with the ability to inactivate SARS-CoV-2 at the time of sample collection could simplify and expand testing for COVID-19 to non-conventional settings.<h4>Methods</h4>We evaluated a guanidium thiocyanate-based buffer, eNAT™ (Copan) as a possible transport and inactivation medium for downstream Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) testing to detect SARS-CoV-2. Inactivation of SARS-CoV-2 USA-WA1/2020 in eNAT and in diluted saliva was studied at different incubation times. The stability of viral RNA in eNAT was also evaluated for up to 7 days at room temperature (28°C), refrigerated conditions (4°C) and at 35°C.<h4>Results</h4>SARS-COV-2 virus spiked directly in eNAT could be inactivated at >5.6 log10 PFU/ml within a minute of incubation. When saliva was diluted 1:1 in eNAT, no cytopathic effect (CPE) on VeroE6 cells was observed, although SARS-CoV-2 RNA could be detected even after 30 min incubation and after two cell culture passages. A 1:2 (saliva:eNAT) dilution abrogated both CPE and detectable viral RNA after as little as 5 min incubation in eNAT. SARS-CoV-2 RNA from virus spiked at 5X the limit of detection remained positive up to 7 days of incubation in all tested conditions.<h4>Conclusion</h4>eNAT and similar guanidinium thiocyanate-based media may be of value for transport, stabilization, and processing of clinical samples for RT-PCR based SARS-CoV-2 detection.Sukalyani BanikKaheerman SaibireShraddha SuryavanshiGlenn JohnsSoumitesh ChakravortyRobert KwiatkowskiDavid AllandPadmapriya P BanadaPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 6, p e0252687 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Sukalyani Banik Kaheerman Saibire Shraddha Suryavanshi Glenn Johns Soumitesh Chakravorty Robert Kwiatkowski David Alland Padmapriya P Banada Inactivation of SARS-CoV-2 virus in saliva using a guanidium based transport medium suitable for RT-PCR diagnostic assays.
description	<h4>Background</h4>Upper respiratory samples used to test for SARS-CoV-2 virus may be infectious and present a hazard during transport and testing. A buffer with the ability to inactivate SARS-CoV-2 at the time of sample collection could simplify and expand testing for COVID-19 to non-conventional settings.<h4>Methods</h4>We evaluated a guanidium thiocyanate-based buffer, eNAT™ (Copan) as a possible transport and inactivation medium for downstream Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) testing to detect SARS-CoV-2. Inactivation of SARS-CoV-2 USA-WA1/2020 in eNAT and in diluted saliva was studied at different incubation times. The stability of viral RNA in eNAT was also evaluated for up to 7 days at room temperature (28°C), refrigerated conditions (4°C) and at 35°C.<h4>Results</h4>SARS-COV-2 virus spiked directly in eNAT could be inactivated at >5.6 log10 PFU/ml within a minute of incubation. When saliva was diluted 1:1 in eNAT, no cytopathic effect (CPE) on VeroE6 cells was observed, although SARS-CoV-2 RNA could be detected even after 30 min incubation and after two cell culture passages. A 1:2 (saliva:eNAT) dilution abrogated both CPE and detectable viral RNA after as little as 5 min incubation in eNAT. SARS-CoV-2 RNA from virus spiked at 5X the limit of detection remained positive up to 7 days of incubation in all tested conditions.<h4>Conclusion</h4>eNAT and similar guanidinium thiocyanate-based media may be of value for transport, stabilization, and processing of clinical samples for RT-PCR based SARS-CoV-2 detection.
format	article
author	Sukalyani Banik Kaheerman Saibire Shraddha Suryavanshi Glenn Johns Soumitesh Chakravorty Robert Kwiatkowski David Alland Padmapriya P Banada
author_facet	Sukalyani Banik Kaheerman Saibire Shraddha Suryavanshi Glenn Johns Soumitesh Chakravorty Robert Kwiatkowski David Alland Padmapriya P Banada
author_sort	Sukalyani Banik
title	Inactivation of SARS-CoV-2 virus in saliva using a guanidium based transport medium suitable for RT-PCR diagnostic assays.
title_short	Inactivation of SARS-CoV-2 virus in saliva using a guanidium based transport medium suitable for RT-PCR diagnostic assays.
title_full	Inactivation of SARS-CoV-2 virus in saliva using a guanidium based transport medium suitable for RT-PCR diagnostic assays.
title_fullStr	Inactivation of SARS-CoV-2 virus in saliva using a guanidium based transport medium suitable for RT-PCR diagnostic assays.
title_full_unstemmed	Inactivation of SARS-CoV-2 virus in saliva using a guanidium based transport medium suitable for RT-PCR diagnostic assays.
title_sort	inactivation of sars-cov-2 virus in saliva using a guanidium based transport medium suitable for rt-pcr diagnostic assays.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/aac5e2b0b2c6472c9b2acee91bd089af
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Inactivation of SARS-CoV-2 virus in saliva using a guanidium based transport medium suitable for RT-PCR diagnostic assays.

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