A simplified SARS-CoV-2 detection protocol for research laboratories.
COVID-19
/ diagnosis
COVID-19 Nucleic Acid Testing
/ methods
COVID-19 Testing
Centers for Disease Control and Prevention, U.S.
HeLa Cells
Humans
Nasopharynx
/ virology
Nucleic Acid Amplification Techniques
/ methods
Oropharynx
/ virology
Pandemics
RNA, Viral
/ isolation & purification
Real-Time Polymerase Chain Reaction
SARS-CoV-2
/ isolation & purification
Saliva
/ virology
Sensitivity and Specificity
United States
Journal
PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081
Informations de publication
Date de publication:
2020
2020
Historique:
received:
25
08
2020
accepted:
07
12
2020
entrez:
18
12
2020
pubmed:
19
12
2020
medline:
5
1
2021
Statut:
epublish
Résumé
Widespread testing is required to limit the current public health crisis caused by the COVID-19 pandemic. Multiple tests protocols have been authorized by the food and drugs administration (FDA) under an emergency use authorization (EUA). The majority of these protocols are based on the gold-standard RT-qPCR test pioneered by the U.S. Centers for Disease Control and Prevention (CDC). However, there is still a widespread lack of testing in the US and many of the clinical diagnostics protocols require extensive human labor and materials that could face supply shortages and present biosafety concerns. Given the need to develop alternative reagents and approaches to provide nucleic-acid testing in the face of heightened demand and potential shortages, we have developed a simplified SARS-CoV-2 testing protocol adapted for its use in research laboratories with minimal molecular biology equipment and expertise. The protocol utilizes TRIzol to purify the viral RNA from different types of clinical specimens, requires minimal BSL-1 precautions and, given its high sensitivity, can be easily adapted to pooling samples strategies.
Identifiants
pubmed: 33338082
doi: 10.1371/journal.pone.0244271
pii: PONE-D-20-26723
pmc: PMC7748277
doi:
Substances chimiques
RNA, Viral
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0244271Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
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