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...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
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Public Library of Science
2023
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| _version_ | 1826310844729786368 |
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| author | Ngetsa, C Osoti, V Okanda, D Marura, F Shah, K Bejon, P Karanja, H Mugo, D Gitonga, J Mutunga, M Lewa, C Orindi, B Ochola-Oyier, LI |
| author_facet | Ngetsa, C Osoti, V Okanda, D Marura, F Shah, K Bejon, P Karanja, H Mugo, D Gitonga, J Mutunga, M Lewa, C Orindi, B Ochola-Oyier, LI |
| author_sort | Ngetsa, C |
| collection | OXFORD |
| 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. |
| first_indexed | 2024-03-07T07:58:02Z |
| format | Journal article |
| id | oxford-uuid:c48225c6-b288-46f6-b10a-1ea02b72efa1 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T07:58:02Z |
| publishDate | 2023 |
| publisher | Public Library of Science |
| record_format | dspace |
| spelling | oxford-uuid:c48225c6-b288-46f6-b10a-1ea02b72efa12023-09-05T14:33:54ZValidation of saline, PBS and a locally produced VTM at varying storage conditions to detect the SARS-CoV-2 virus by qRT-PCRJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:c48225c6-b288-46f6-b10a-1ea02b72efa1EnglishSymplectic ElementsPublic Library of Science2023Ngetsa, COsoti, VOkanda, DMarura, FShah, KBejon, PKaranja, HMugo, DGitonga, JMutunga, MLewa, COrindi, BOchola-Oyier, LICoronavirus 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. |
| spellingShingle | Ngetsa, C Osoti, V Okanda, D Marura, F Shah, K Bejon, P Karanja, H Mugo, D Gitonga, J Mutunga, M Lewa, C Orindi, B Ochola-Oyier, LI 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 |
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