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Validation of saline, PBS and a locally produced VTM at varying storage conditions to detect the SARS-CoV-2 virus by qRT-PCR

Coronavirus Disease-2019 tests require a Nasopharyngeal (NP) and/or Oropharyngeal (OP) specimen from the upper airway, from which virus RNA is extracted and detected through quantitative reverse transcription-Polymerase Chain Reaction (qRT-PCR). The viability of the virus is maintained after collect...

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Autores principales: Ngetsa, Caroline, Osoti, Victor, Okanda, Dorcas, Marura, Faith, Shah, Krupali, Karanja, Henry, Mugo, Daisy, Gitonga, John, Mutunga, Martin, Lewa, Clement, Orindi, Benedict, Bejon, Philip, Ochola-Oyier, Lynette Isabella
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9924993/
https://www.ncbi.nlm.nih.gov/pubmed/36780469
http://dx.doi.org/10.1371/journal.pone.0280685
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author Ngetsa, Caroline
Osoti, Victor
Okanda, Dorcas
Marura, Faith
Shah, Krupali
Karanja, Henry
Mugo, Daisy
Gitonga, John
Mutunga, Martin
Lewa, Clement
Orindi, Benedict
Bejon, Philip
Ochola-Oyier, Lynette Isabella
author_facet Ngetsa, Caroline
Osoti, Victor
Okanda, Dorcas
Marura, Faith
Shah, Krupali
Karanja, Henry
Mugo, Daisy
Gitonga, John
Mutunga, Martin
Lewa, Clement
Orindi, Benedict
Bejon, Philip
Ochola-Oyier, Lynette Isabella
author_sort Ngetsa, Caroline
collection PubMed
description Coronavirus Disease-2019 tests require a Nasopharyngeal (NP) and/or Oropharyngeal (OP) specimen from the upper airway, from which virus RNA is extracted and detected through quantitative reverse transcription-Polymerase Chain Reaction (qRT-PCR). The viability of the virus is maintained after collection by storing the NP/OP swabs in Viral Transport Media (VTM). We evaluated the performance of four transport media: locally manufactured (“REVITAL”) Viral Transport Media (RVTM), Standard Universal Transport Media (SUTM), PBS and 0.9% (w/v) NaCl (normal saline). We used laboratory cultured virus to evaluate: i) viral recovery and maintaining integrity at different time periods and temperatures; ii) stability in yielding detectable RNA consistently for all time points and conditions; and iii) their overall accuracy. Four vials of SARS-CoV-2 cultured virus (2 high and 2 low concentration samples) and 1 negative control sample were prepared for each media type (SUTM, RVTM, PBS and normal saline) and stored at the following temperatures, -80°C, 4°C, 25°C and 37°C for 7 days. Viral RNA extractions and qRT-PCR were performed at 1, 2, 3, 4 and 7 days after inoculation with the cultured virus to assess virus stability and viral recovery. Ct values fell over time at 25°C and 37°C, but normal saline, PBS, RVTM and SUTM all showed comparable performance in maintaining virus integrity and stability allowing for the detection of SARS-CoV-2 RNA. Overall, this study demonstrated that normal saline, PBS and the locally manufactured VTM can be used for COVID-19 sample collection and testing, thus expanding the range of SARS-CoV-2 viral collection media.
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spelling pubmed-99249932023-02-14 Validation of saline, PBS and a locally produced VTM at varying storage conditions to detect the SARS-CoV-2 virus by qRT-PCR Ngetsa, Caroline Osoti, Victor Okanda, Dorcas Marura, Faith Shah, Krupali Karanja, Henry Mugo, Daisy Gitonga, John Mutunga, Martin Lewa, Clement Orindi, Benedict Bejon, Philip Ochola-Oyier, Lynette Isabella PLoS One Research Article Coronavirus Disease-2019 tests require a Nasopharyngeal (NP) and/or Oropharyngeal (OP) specimen from the upper airway, from which virus RNA is extracted and detected through quantitative reverse transcription-Polymerase Chain Reaction (qRT-PCR). The viability of the virus is maintained after collection by storing the NP/OP swabs in Viral Transport Media (VTM). We evaluated the performance of four transport media: locally manufactured (“REVITAL”) Viral Transport Media (RVTM), Standard Universal Transport Media (SUTM), PBS and 0.9% (w/v) NaCl (normal saline). We used laboratory cultured virus to evaluate: i) viral recovery and maintaining integrity at different time periods and temperatures; ii) stability in yielding detectable RNA consistently for all time points and conditions; and iii) their overall accuracy. Four vials of SARS-CoV-2 cultured virus (2 high and 2 low concentration samples) and 1 negative control sample were prepared for each media type (SUTM, RVTM, PBS and normal saline) and stored at the following temperatures, -80°C, 4°C, 25°C and 37°C for 7 days. Viral RNA extractions and qRT-PCR were performed at 1, 2, 3, 4 and 7 days after inoculation with the cultured virus to assess virus stability and viral recovery. Ct values fell over time at 25°C and 37°C, but normal saline, PBS, RVTM and SUTM all showed comparable performance in maintaining virus integrity and stability allowing for the detection of SARS-CoV-2 RNA. Overall, this study demonstrated that normal saline, PBS and the locally manufactured VTM can be used for COVID-19 sample collection and testing, thus expanding the range of SARS-CoV-2 viral collection media. Public Library of Science 2023-02-13 /pmc/articles/PMC9924993/ /pubmed/36780469 http://dx.doi.org/10.1371/journal.pone.0280685 Text en © 2023 Ngetsa et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://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
Ngetsa, Caroline
Osoti, Victor
Okanda, Dorcas
Marura, Faith
Shah, Krupali
Karanja, Henry
Mugo, Daisy
Gitonga, John
Mutunga, Martin
Lewa, Clement
Orindi, Benedict
Bejon, Philip
Ochola-Oyier, Lynette Isabella
Validation of saline, PBS and a locally produced VTM at varying storage conditions to detect the SARS-CoV-2 virus by qRT-PCR
title Validation of saline, PBS and a locally produced VTM at varying storage conditions to detect the SARS-CoV-2 virus by qRT-PCR
title_full Validation of saline, PBS and a locally produced VTM at varying storage conditions to detect the SARS-CoV-2 virus by qRT-PCR
title_fullStr Validation of saline, PBS and a locally produced VTM at varying storage conditions to detect the SARS-CoV-2 virus by qRT-PCR
title_full_unstemmed Validation of saline, PBS and a locally produced VTM at varying storage conditions to detect the SARS-CoV-2 virus by qRT-PCR
title_short Validation of saline, PBS and a locally produced VTM at varying storage conditions to detect the SARS-CoV-2 virus by qRT-PCR
title_sort validation of saline, pbs and a locally produced vtm at varying storage conditions to detect the sars-cov-2 virus by qrt-pcr
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9924993/
https://www.ncbi.nlm.nih.gov/pubmed/36780469
http://dx.doi.org/10.1371/journal.pone.0280685
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