Preparation of the luciferase-labeled antibody for improving the detection sensitivity of viral antigen.
Antigen detection
Automatic magnet chemiluminescence immune assay (AMCA)
Highly sensitive
Nano luciferase
SARS-CoV-2
Journal
Virology journal
ISSN: 1743-422X
Titre abrégé: Virol J
Pays: England
ID NLM: 101231645
Informations de publication
Date de publication:
28 07 2022
28 07 2022
Historique:
received:
23
05
2022
accepted:
17
07
2022
entrez:
28
7
2022
pubmed:
29
7
2022
medline:
2
8
2022
Statut:
epublish
Résumé
Viral antigen detection test is the most common method used to detect viruses in the field rapidly. However, due to the low sensitivity, it can only be used as an auxiliary diagnosis method for virus infection. Improving sensitivity is crucial for developing more accurate viral antigen tests. Nano luciferase (Nluc) is a sensitive reporter that has not been used in virus detection. In this study, we produced an intracellularly Nluc labeled detection antibody (Nluc-ch2C5) and evaluated its ability to improve the detection sensitivity of respiratory syndrome coronavirus 2 (SARS-CoV-2) antigens. Compared with the traditional horse-radish peroxidase (HRP) labeled antibody (HRP-ch2C5), Nluc-ch2C5 was 41 times more sensitive for inactivated SARS-CoV-2 virus by sandwich chemiluminescence ELISA. Then we applied Nluc-ch2C5 to establish an automatic magnet chemiluminescence immune assay (AMCA) for the SARS-CoV-2 viral spike protein, the limit of detection was 68 pfu/reaction. The clinical sensitivity and specificity reached 75% (24/32) and 100% (48/48) using 32 PCR-positive and 48 PCR-negative swab samples for clinical evaluation, which is more sensitive than the commercial ELSA kit and colloid gold strip kit. Here, monoclonal antibody ch2C5 served as a model antibody and the SARS-CoV-2 served as a model pathogen. The Nluc labeled detecting antibody (Nluc-ch2C5) significantly improved the detection sensitivity of SARS-CoV-2 antigen. This labeling principle applies to other viral infections, so this labeling and test format could be expected to play an important role in detecting other virus antigens.
Sections du résumé
BACKGROUND
Viral antigen detection test is the most common method used to detect viruses in the field rapidly. However, due to the low sensitivity, it can only be used as an auxiliary diagnosis method for virus infection. Improving sensitivity is crucial for developing more accurate viral antigen tests. Nano luciferase (Nluc) is a sensitive reporter that has not been used in virus detection.
RESULTS
In this study, we produced an intracellularly Nluc labeled detection antibody (Nluc-ch2C5) and evaluated its ability to improve the detection sensitivity of respiratory syndrome coronavirus 2 (SARS-CoV-2) antigens. Compared with the traditional horse-radish peroxidase (HRP) labeled antibody (HRP-ch2C5), Nluc-ch2C5 was 41 times more sensitive for inactivated SARS-CoV-2 virus by sandwich chemiluminescence ELISA. Then we applied Nluc-ch2C5 to establish an automatic magnet chemiluminescence immune assay (AMCA) for the SARS-CoV-2 viral spike protein, the limit of detection was 68 pfu/reaction. The clinical sensitivity and specificity reached 75% (24/32) and 100% (48/48) using 32 PCR-positive and 48 PCR-negative swab samples for clinical evaluation, which is more sensitive than the commercial ELSA kit and colloid gold strip kit.
CONCLUSIONS
Here, monoclonal antibody ch2C5 served as a model antibody and the SARS-CoV-2 served as a model pathogen. The Nluc labeled detecting antibody (Nluc-ch2C5) significantly improved the detection sensitivity of SARS-CoV-2 antigen. This labeling principle applies to other viral infections, so this labeling and test format could be expected to play an important role in detecting other virus antigens.
Identifiants
pubmed: 35902865
doi: 10.1186/s12985-022-01855-6
pii: 10.1186/s12985-022-01855-6
pmc: PMC9332066
doi:
Substances chimiques
Antigens, Viral
0
Luciferases
EC 1.13.12.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
126Informations de copyright
© 2022. The Author(s).
Références
Biosens Bioelectron. 2021 Jan 1;171:112686
pubmed: 33086175
Lancet. 2020 Feb 15;395(10223):470-473
pubmed: 31986257
Vector Borne Zoonotic Dis. 2019 May;19(5):365-369
pubmed: 30431406
J Immunol Methods. 2012 Apr 30;378(1-2):102-15
pubmed: 22370429
Curr Opin Struct Biol. 2014 Feb;24:1-9
pubmed: 24721448
Front Immunol. 2021 Apr 23;12:670626
pubmed: 33968077
J Clin Virol. 2021 Jan;134:104712
pubmed: 33338894
J Zhejiang Univ Sci B. 2020 May;21(5):343-360
pubmed: 32425000
Protein Expr Purif. 2018 Sep;149:75-83
pubmed: 29655788
Cold Spring Harb Protoc. 2019 Mar 1;2019(3):
pubmed: 30824621
Virol Sin. 2020 Dec;35(6):699-712
pubmed: 33351166
ACS Chem Biol. 2012 Nov 16;7(11):1848-57
pubmed: 22894855
MAbs. 2015;7(3):505-15
pubmed: 25759214
Virol Sin. 2021 Dec;36(6):1484-1491
pubmed: 34403037
Food Chem. 2019 Mar 1;275:721-729
pubmed: 30724255
PLoS Negl Trop Dis. 2020 Oct 28;14(10):e0008856
pubmed: 33112863
Clin Diagn Virol. 1996 May;5(2-3):81-90
pubmed: 15566866
Virol J. 2022 Mar 28;19(1):54
pubmed: 35346227
Methods Mol Biol. 2021;2247:59-76
pubmed: 33301112
Emerg Microbes Infect. 2020 Dec;9(1):1489-1496
pubmed: 32543298
Cold Spring Harb Protoc. 2020 Jul 1;2020(7):099242
pubmed: 32611784
Cold Spring Harb Protoc. 2019 Mar 1;2019(3):
pubmed: 30824622
MAbs. 2021 Jan-Dec;13(1):1953683
pubmed: 34313527
mBio. 2021 May 18;12(3):
pubmed: 34006662
Virol J. 2021 Jun 29;18(1):131
pubmed: 34187522
Virol Sin. 2022 Feb;37(1):138-141
pubmed: 35234627
Science. 2010 May 7;328(5979):704-8
pubmed: 20448176
Lancet. 2019 Mar 2;393(10174):936-948
pubmed: 30777297