Rapid and accurate nucleobase detection using FnCas9 and its application in COVID-19 diagnosis.
CRISPRDx
FELUDA
FnCas9
LFA
SARS-CoV2
SNV detection
Journal
Biosensors & bioelectronics
ISSN: 1873-4235
Titre abrégé: Biosens Bioelectron
Pays: England
ID NLM: 9001289
Informations de publication
Date de publication:
01 Jul 2021
01 Jul 2021
Historique:
received:
05
03
2021
revised:
22
03
2021
accepted:
26
03
2021
pubmed:
19
4
2021
medline:
15
5
2021
entrez:
18
4
2021
Statut:
ppublish
Résumé
Rapid detection of DNA/RNA pathogenic sequences or variants through point-of-care diagnostics is valuable for accelerated clinical prognosis, as witnessed during the recent COVID-19 outbreak. Traditional methods relying on qPCR or sequencing are tough to implement with limited resources, necessitating the development of accurate and robust alternative strategies. Here, we report FnCas9 Editor Linked Uniform Detection Assay (FELUDA) that utilizes a direct Cas9 based enzymatic readout for detecting nucleobase and nucleotide sequences without trans-cleavage of reporter molecules. We also demonstrate that FELUDA is 100% accurate in detecting single nucleotide variants (SNVs), including heterozygous carriers, and present a simple web-tool JATAYU to aid end-users. FELUDA is semi-quantitative, can adapt to multiple signal detection platforms, and deploy for versatile applications such as molecular diagnosis during infectious disease outbreaks like COVID-19. Employing a lateral flow readout, FELUDA shows 100% sensitivity and 97% specificity across all ranges of viral loads in clinical samples within 1hr. In combination with RT-RPA and a smartphone application True Outcome Predicted via Strip Evaluation (TOPSE), we present a prototype for FELUDA for CoV-2 detection closer to home.
Identifiants
pubmed: 33866136
pii: S0956-5663(21)00244-X
doi: 10.1016/j.bios.2021.113207
pmc: PMC8020606
pii:
doi:
Substances chimiques
RNA, Viral
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
113207Informations de copyright
Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.
Références
Trends Biotechnol. 2019 Jul;37(7):730-743
pubmed: 30654914
Science. 2018 Apr 27;360(6387):439-444
pubmed: 29449508
Ann Clin Microbiol Antimicrob. 2003 Nov 21;2:11
pubmed: 14633281
Science. 2018 Apr 27;360(6387):381-382
pubmed: 29700254
ACS Synth Biol. 2018 Mar 16;7(3):807-813
pubmed: 29486117
Int J Food Sci Nutr. 2018 Mar;69(2):245-252
pubmed: 28738701
Nature. 2017 Oct 19;550(7676):407-410
pubmed: 28931002
Genome Biol. 2019 Jul 1;20(1):132
pubmed: 31262344
Bioinformatics. 2010 Mar 15;26(6):841-2
pubmed: 20110278
J Transl Med. 2020 Apr 22;18(1):179
pubmed: 32321524
PLoS One. 2021 Feb 17;16(2):e0247115
pubmed: 33596239
Cell Res. 2018 Apr;28(4):491-493
pubmed: 29531313
Genome Med. 2018 Apr 24;10(1):32
pubmed: 29690921
Mil Med Res. 2020 Mar 13;7(1):11
pubmed: 32169119
Science. 2018 Apr 27;360(6387):436-439
pubmed: 29449511
J Mol Biol. 2003 Aug 29;331(5):991-1004
pubmed: 12927536
Nucleic Acids Res. 2019 Aug 22;47(14):e83
pubmed: 31114866
Science. 2016 Aug 5;353(6299):aaf5573
pubmed: 27256883
ACS Synth Biol. 2019 Oct 18;8(10):2228-2237
pubmed: 31532637
Cell. 2020 May 14;181(4):914-921.e10
pubmed: 32330414
Bioinformatics. 2010 Mar 1;26(5):589-95
pubmed: 20080505
Cell. 2016 May 19;165(5):1255-1266
pubmed: 27160350
Nat Biotechnol. 2020 Jul;38(7):870-874
pubmed: 32300245
Proc Natl Acad Sci U S A. 2019 Oct 15;116(42):20959-20968
pubmed: 31570623
Nat Protoc. 2019 Oct;14(10):2986-3012
pubmed: 31548639
N Engl J Med. 2017 Apr 20;376(16):1561-1573
pubmed: 28423290
Mol Cell. 2018 Apr 19;70(2):327-339.e5
pubmed: 29551514
Cell. 2016 Feb 25;164(5):950-61
pubmed: 26875867
Mol Cell. 2017 Feb 16;65(4):618-630.e7
pubmed: 28065598
Curr Opin Hematol. 2020 May;27(3):141-148
pubmed: 32167946
Bioinformatics. 2008 Oct 15;24(20):2395-6
pubmed: 18697769
J Clin Microbiol. 2020 Apr 23;58(5):
pubmed: 32132196
PLoS One. 2013 Aug 01;8(8):e70266
pubmed: 23936402
Euro Surveill. 2017 Mar 30;22(13):
pubmed: 28382917
Lancet. 2020 Apr 25;395(10233):1321-1322
pubmed: 32277876
Clin Biochem Rev. 2006 Feb;27(1):63-75
pubmed: 16886048
Chem Rev. 2015 Nov 25;115(22):12491-545
pubmed: 26551336
Front Microbiol. 2018 Dec 18;9:3154
pubmed: 30619218
Trends Analyt Chem. 2018 Jan;98:19-35
pubmed: 32287544
Nat Biomed Eng. 2020 Dec;4(12):1140-1149
pubmed: 32848209
Lancet. 2020 Apr 4;395(10230):1101-1102
pubmed: 32247384
Nucleic Acids Res. 2015 Jan;43(Database issue):D571-7
pubmed: 25428358
Lancet Infect Dis. 2020 Jun;20(6):631-633
pubmed: 32213329
Nucleic Acids Res. 2014 Jan;42(Database issue):D980-5
pubmed: 24234437
Nature. 2020 Mar;579(7798):265-269
pubmed: 32015508
Am J Clin Pathol. 2020 May 5;153(6):706-708
pubmed: 32227199