A Nuclease Protection ELISA Assay for Colorimetric and Electrochemical Detection of Nucleic Acids.
Journal
Analytical methods : advancing methods and applications
ISSN: 1759-9660
Titre abrégé: Anal Methods
Pays: England
ID NLM: 101519733
Informations de publication
Date de publication:
28 Feb 2019
28 Feb 2019
Historique:
entrez:
29
10
2019
pubmed:
28
10
2019
medline:
28
10
2019
Statut:
ppublish
Résumé
Early and accurate diagnosis is crucial to monitor infection outcomes and provide timely interventions. However, gold standard polymerase chain reaction assays (PCR) are labor-intensive and require expensive reagents and instrumentation. Nuclease protection has been used for decades to detect and quantify nucleic acid but has not yet been investigated as a diagnostic tool for infectious disease. In this work, we describe a nuclease protection enzyme-linked immunosorbent assay (NP-ELISA) for accurate and sensitive detection of nucleic acid. Briefly, binding of a nucleic acid target to an oligo probe protects it from digestion of un-hybridized nucleic acid by S1 nuclease. Following the workflow of an ELISA, a horseradish peroxidase (HRP)-conjugated antibody binds the probe and oxidizes its substrate to generate signal. The assay was validated with three HRP substrates for absorbance, chemiluminescence, and electrochemical readouts, demonstrating great versatility. Electrochemical detection with 3,3',5,5'-Tetramethylbenzidine (TMB) gave the highest assay sensitivity with a limit of detection of 3.72×10
Identifiants
pubmed: 31656535
doi: 10.1039/C8AY02729C
pmc: PMC6814143
mid: NIHMS1008985
doi:
Types de publication
Journal Article
Langues
eng
Pagination
1027-1034Subventions
Organisme : NIAID NIH HHS
ID : R01 AI114675
Pays : United States
Organisme : NIAID NIH HHS
ID : R01 AI132668
Pays : United States
Déclaration de conflit d'intérêts
Conflicts of interest We have no conflicts to declare.
Références
Diagn Microbiol Infect Dis. 2018 May;91(1):13-19
pubmed: 29398460
Biosens Bioelectron. 2016 Mar 15;77:457-63
pubmed: 26454827
Mar Biotechnol (NY). 2012 Aug;14(4):502-11
pubmed: 22282220
Mol Biotechnol. 2012 Sep;52(1):8-15
pubmed: 22072316
Cell Res. 2014 Jun;24(6):766-9
pubmed: 24710597
Neth J Med. 2016 Mar;74(3):104-9
pubmed: 27020989
J Nanobiotechnology. 2011 Mar 17;9:8
pubmed: 21414201
Anal Chem. 2018 Jun 19;90(12):7777-7783
pubmed: 29790331
Anal Sci. 2016;32(8):815-8
pubmed: 27506705
Anal Chim Acta. 2015 Sep 3;891:247-54
pubmed: 26388383
Lab Invest. 2007 Oct;87(10):979-97
pubmed: 17700562
Biosens Bioelectron. 2016 May 15;79:121-9
pubmed: 26703990
Gynecol Oncol. 1992 Nov;47(2):228-33
pubmed: 1361480
Clin Chim Acta. 2015 Apr 15;444:199-205
pubmed: 25689793
J Virol. 2007 Dec;81(24):13771-82
pubmed: 17942535
Methods Mol Biol. 2011;721:173-82
pubmed: 21431685
J Biol Chem. 1982 Apr 10;257(7):3669-75
pubmed: 6277943
Anal Chem. 1985 Oct;57(12):1321A-1331A
pubmed: 4061832
Biotechnol Lett. 2016 Jan;38(1):57-63
pubmed: 26334938
Methods Enzymol. 2013;530:89-97
pubmed: 24034316
Talanta. 2012 Sep 15;99:375-9
pubmed: 22967567
Cell. 1977 Nov;12(3):721-32
pubmed: 922889
Hum Mutat. 2005 Jun;25(6):575-82
pubmed: 15880407
Am J Orthod Dentofacial Orthop. 2014 Jan;145(1):36-40
pubmed: 24373653
Clin Microbiol Rev. 2016 Jul;29(3):487-524
pubmed: 27029595
Talanta. 2012 Jan 15;88:468-76
pubmed: 22265528
Nucleic Acids Res. 1998 Oct 15;26(20):4597-602
pubmed: 9753726
Nucleic Acids Res. 2004 May 11;32(8):2632-41
pubmed: 15141034
Biosens Bioelectron. 2017 Feb 15;88:9-14
pubmed: 27297187
Nature. 2007 Oct 11;449(7163):745-7
pubmed: 17898712
Curr Genomics. 2016 Dec;17(6):499-508
pubmed: 28217006
Anal Biochem. 1996 Apr 5;236(1):182-4
pubmed: 8619489
J Phys Chem B. 2009 Oct 22;113(42):14021-5
pubmed: 19780515
Toxicol Appl Pharmacol. 2015 Jun 15;285(3):198-206
pubmed: 25818602
Angew Chem Int Ed Engl. 2012 Jan 23;51(4):1019-23
pubmed: 22173871