A Generic, Scalable, and Rapid Time-Resolved Förster Resonance Energy Transfer-Based Assay for Antigen Detection-SARS-CoV-2 as a Proof of Concept.
Antigens, Viral
/ analysis
COVID-19
/ diagnosis
COVID-19 Serological Testing
/ methods
Coronavirus Nucleocapsid Proteins
/ analysis
Fluorescence Resonance Energy Transfer
Humans
Limit of Detection
Nasopharynx
/ virology
Phosphoproteins
/ analysis
Proof of Concept Study
Recombinant Proteins
/ immunology
SARS-CoV-2
/ immunology
Sensitivity and Specificity
Spike Glycoprotein, Coronavirus
/ analysis
Time Factors
COVID-19
SARS-CoV-2
TR-FRET
antigen test
mix and read
rapid diagnostic test
Journal
mBio
ISSN: 2150-7511
Titre abrégé: mBio
Pays: United States
ID NLM: 101519231
Informations de publication
Date de publication:
18 05 2021
18 05 2021
Historique:
entrez:
19
5
2021
pubmed:
20
5
2021
medline:
2
6
2021
Statut:
epublish
Résumé
The ongoing coronavirus disease 2019 (COVID-19) pandemic has seen an unprecedented increase in the demand for rapid and reliable diagnostic tools, leaving many laboratories scrambling for resources. We present a fast and simple assay principle for antigen detection and demonstrate its functionality by detecting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antigens in nasopharyngeal swabs. The method is based on the detection of SARS-CoV-2 nucleoprotein (NP) and S protein (SP) via time-resolved Förster resonance energy transfer (TR-FRET) with donor- and acceptor-labeled polyclonal anti-NP and -SP antibodies. Using recombinant proteins and cell culture-grown SARS-CoV-2, the limits of detection were established as 25 pg of NP or 20 infectious units (IU) and 875 pg of SP or 625 IU. Testing reverse transcription-PCR (RT-PCR)-positive (
Identifiants
pubmed: 34006662
pii: mBio.00902-21
doi: 10.1128/mBio.00902-21
pmc: PMC8262888
pii:
doi:
Substances chimiques
Antigens, Viral
0
Coronavirus Nucleocapsid Proteins
0
Phosphoproteins
0
Recombinant Proteins
0
Spike Glycoprotein, Coronavirus
0
nucleocapsid phosphoprotein, SARS-CoV-2
0
spike protein, SARS-CoV-2
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Informations de copyright
Copyright © 2021 Rusanen et al.
Références
J Clin Microbiol. 2015 Jul;53(7):2292-7
pubmed: 25972427
Euro Surveill. 2020 Jan;25(3):
pubmed: 31992387
PLoS One. 2013 May 07;8(5):e62739
pubmed: 23667515
Clin Infect Dis. 2020 Dec 17;71(10):2663-2666
pubmed: 32442256
Proc Natl Acad Sci U S A. 2005 May 31;102(22):7988-93
pubmed: 15897467
Nature. 2020 May;581(7809):465-469
pubmed: 32235945
PLoS One. 2019 Jul 23;14(7):e0219474
pubmed: 31335898
PLoS One. 2019 Nov 26;14(11):e0225851
pubmed: 31770411
Euro Surveill. 2020 Mar;25(11):
pubmed: 32209163
Cochrane Database Syst Rev. 2020 Aug 26;8:CD013705
pubmed: 32845525
PLoS One. 2014 Sep 02;9(9):e106432
pubmed: 25181527
J Mol Diagn. 2021 Apr;23(4):407-416
pubmed: 33486074
J Virol. 2004 Nov;78(21):11739-50
pubmed: 15479815
Nat Med. 2020 Jul;26(7):1033-1036
pubmed: 32398876
Viruses. 2021 Jan 20;13(2):
pubmed: 33498157
J Virol. 2015 Feb;89(4):1954-64
pubmed: 25428871
Curr Protoc Microbiol. 2020 Jun;57(1):e100
pubmed: 32302069
Clin Infect Dis. 2021 Nov 2;73(9):e2861-e2866
pubmed: 33479756
J Infect Dis. 2020 Oct 1;222(9):1462-1467
pubmed: 32798217
N Engl J Med. 2020 Nov 26;383(22):e120
pubmed: 32997903
N Engl J Med. 2020 May 28;382(22):2081-2090
pubmed: 32329971
J Clin Microbiol. 2015 Feb;53(2):636-40
pubmed: 25520445
J Clin Microbiol. 2020 Oct 21;58(11):
pubmed: 32839250
Cell. 2020 Apr 16;181(2):271-280.e8
pubmed: 32142651