tetano
Editor, Senior Moderator
PLoS One
. 2023 Feb 13;18(2):e0280685.
doi: 10.1371/journal.pone.0280685. eCollection 2023.
Validation of saline, PBS and a locally produced VTM at varying storage conditions to detect the SARS-CoV-2 virus by qRT-PCR
Caroline Ngetsa[SUP] 1 [/SUP], Victor Osoti[SUP] 1 [/SUP], Dorcas Okanda[SUP] 1 [/SUP], Faith Marura[SUP] 1 [/SUP], Krupali Shah[SUP] 2 [/SUP], Henry Karanja[SUP] 1 [/SUP], Daisy Mugo[SUP] 1 [/SUP], John Gitonga[SUP] 1 [/SUP], Martin Mutunga[SUP] 1 [/SUP], Clement Lewa[SUP] 1 [/SUP], Benedict Orindi[SUP] 1 [/SUP], Philip Bejon[SUP] 1 3 [/SUP], Lynette Isabella Ochola-Oyier[SUP] 1 [/SUP]
Affiliations
Abstract
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.
. 2023 Feb 13;18(2):e0280685.
doi: 10.1371/journal.pone.0280685. eCollection 2023.
Validation of saline, PBS and a locally produced VTM at varying storage conditions to detect the SARS-CoV-2 virus by qRT-PCR
Caroline Ngetsa[SUP] 1 [/SUP], Victor Osoti[SUP] 1 [/SUP], Dorcas Okanda[SUP] 1 [/SUP], Faith Marura[SUP] 1 [/SUP], Krupali Shah[SUP] 2 [/SUP], Henry Karanja[SUP] 1 [/SUP], Daisy Mugo[SUP] 1 [/SUP], John Gitonga[SUP] 1 [/SUP], Martin Mutunga[SUP] 1 [/SUP], Clement Lewa[SUP] 1 [/SUP], Benedict Orindi[SUP] 1 [/SUP], Philip Bejon[SUP] 1 3 [/SUP], Lynette Isabella Ochola-Oyier[SUP] 1 [/SUP]
Affiliations
- PMID: 36780469
- DOI: 10.1371/journal.pone.0280685
Abstract
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.