PMID- 36780469
OWN - NLM
STAT- MEDLINE
DCOM- 20230215
LR  - 20240422
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 18
IP  - 2
DP  - 2023
TI  - Validation of saline, PBS and a locally produced VTM at varying storage 
      conditions to detect the SARS-CoV-2 virus by qRT-PCR.
PG  - e0280685
LID - 10.1371/journal.pone.0280685 [doi]
LID - e0280685
AB  - 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 degrees C, 4 degrees C, 25 degrees C and 37 degrees 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 degrees C and 37 degrees 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.
CI  - Copyright: (c) 2023 Ngetsa et al. This is an open access article distributed under 
      the terms of the Creative Commons Attribution License, which permits unrestricted 
      use, distribution, and reproduction in any medium, provided the original author 
      and source are credited.
FAU - Ngetsa, Caroline
AU  - Ngetsa C
AD  - KEMRI-Wellcome Trust Research Programme, Centre for Geographic Medicine Research 
      Coast, Kilifi, Kenya.
FAU - Osoti, Victor
AU  - Osoti V
AD  - KEMRI-Wellcome Trust Research Programme, Centre for Geographic Medicine Research 
      Coast, Kilifi, Kenya.
FAU - Okanda, Dorcas
AU  - Okanda D
AD  - KEMRI-Wellcome Trust Research Programme, Centre for Geographic Medicine Research 
      Coast, Kilifi, Kenya.
FAU - Marura, Faith
AU  - Marura F
AD  - KEMRI-Wellcome Trust Research Programme, Centre for Geographic Medicine Research 
      Coast, Kilifi, Kenya.
FAU - Shah, Krupali
AU  - Shah K
AUID- ORCID: 0000-0003-4787-7259
AD  - Revital Healthcare (EPZ) Limited, Mombasa, Kenya.
FAU - Karanja, Henry
AU  - Karanja H
AD  - KEMRI-Wellcome Trust Research Programme, Centre for Geographic Medicine Research 
      Coast, Kilifi, Kenya.
FAU - Mugo, Daisy
AU  - Mugo D
AD  - KEMRI-Wellcome Trust Research Programme, Centre for Geographic Medicine Research 
      Coast, Kilifi, Kenya.
FAU - Gitonga, John
AU  - Gitonga J
AD  - KEMRI-Wellcome Trust Research Programme, Centre for Geographic Medicine Research 
      Coast, Kilifi, Kenya.
FAU - Mutunga, Martin
AU  - Mutunga M
AD  - KEMRI-Wellcome Trust Research Programme, Centre for Geographic Medicine Research 
      Coast, Kilifi, Kenya.
FAU - Lewa, Clement
AU  - Lewa C
AD  - KEMRI-Wellcome Trust Research Programme, Centre for Geographic Medicine Research 
      Coast, Kilifi, Kenya.
FAU - Orindi, Benedict
AU  - Orindi B
AD  - KEMRI-Wellcome Trust Research Programme, Centre for Geographic Medicine Research 
      Coast, Kilifi, Kenya.
FAU - Bejon, Philip
AU  - Bejon P
AD  - KEMRI-Wellcome Trust Research Programme, Centre for Geographic Medicine Research 
      Coast, Kilifi, Kenya.
AD  - Nuffield Department of Medicine, Centre for Clinical Vaccinology and Tropical 
      Medicine, Churchill Hospital, University of Oxford, Oxford, United Kingdom.
FAU - Ochola-Oyier, Lynette Isabella
AU  - Ochola-Oyier LI
AUID- ORCID: 0000-0003-4393-0470
AD  - KEMRI-Wellcome Trust Research Programme, Centre for Geographic Medicine Research 
      Coast, Kilifi, Kenya.
LA  - eng
GR  - WT_/Wellcome Trust/United Kingdom
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20230213
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Saline Solution)
RN  - 0 (RNA, Viral)
SB  - IM
MH  - Humans
MH  - *SARS-CoV-2/genetics
MH  - *COVID-19/diagnosis
MH  - Saline Solution
MH  - RNA, Viral/genetics/analysis
MH  - Specimen Handling
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - COVID-19 Testing
PMC - PMC9924993
COIS- The authors have declared that no competing interests exist.
EDAT- 2023/02/14 06:00
MHDA- 2023/02/16 06:00
PMCR- 2023/02/13
CRDT- 2023/02/13 14:53
PHST- 2022/09/19 00:00 [received]
PHST- 2023/01/05 00:00 [accepted]
PHST- 2023/02/13 14:53 [entrez]
PHST- 2023/02/14 06:00 [pubmed]
PHST- 2023/02/16 06:00 [medline]
PHST- 2023/02/13 00:00 [pmc-release]
AID - PONE-D-22-25994 [pii]
AID - 10.1371/journal.pone.0280685 [doi]
PST - epublish
SO  - PLoS One. 2023 Feb 13;18(2):e0280685. doi: 10.1371/journal.pone.0280685. 
      eCollection 2023.