Large scale production and characterization of SARS-CoV-2 whole antigen for serological test development.
Animals
Antibodies, Viral
/ blood
Antigens, Viral
/ chemistry
Blotting, Western
COVID-19
/ diagnosis
COVID-19 Serological Testing
/ methods
Chlorocebus aethiops
Coronavirus Nucleocapsid Proteins
/ chemistry
Enzyme-Linked Immunosorbent Assay
/ methods
Humans
Phosphoproteins
/ chemistry
SARS-CoV-2
/ chemistry
Spike Glycoprotein, Coronavirus
/ chemistry
Vero Cells
Virus Cultivation
/ methods
COVID-19
ELISA
native antigen
serological test
viral culture
Journal
Journal of clinical laboratory analysis
ISSN: 1098-2825
Titre abrégé: J Clin Lab Anal
Pays: United States
ID NLM: 8801384
Informations de publication
Date de publication:
Apr 2021
Apr 2021
Historique:
revised:
15
01
2021
received:
16
12
2020
accepted:
19
01
2021
pubmed:
21
2
2021
medline:
29
4
2021
entrez:
20
2
2021
Statut:
ppublish
Résumé
The rapid spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has generated a pandemic with alarming rates of fatality worldwide. This situation has had a major impact on clinical laboratories that have attempted to answer the urgent need for diagnostic tools, since the identification of coronavirus disease 2019 (COVID-19). Development of a reliable serological diagnostic immunoassay, with high levels of sensitivity and specificity to detect SARS-CoV-2 antibodies with improved differential diagnosis from other circulating viruses, is mandatory. An enzyme-linked immunosorbent assay (ELISA) using whole inactivated virus cultured in vitro, was developed to detect viral antigens. WB and ELISA investigations were carried out with sera of convalescent patients and negative sera samples. Both analyses were concurrently performed with recombinant MABs to verify the findings. Preliminary data from 10 sera (5 patients with COVID-19, and 5 healthy controls) using this immunoassay are very promising, successfully identifying all of the confirmed SARS-CoV-2-positive individuals. This ELISA appears to be a specific and reliable method for detecting COVID-19 antibodies (IgG, IgM, and IgA), and a useful tool for identifying individuals which have developed immunity to the virus.
Sections du résumé
BACKGROUND
BACKGROUND
The rapid spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has generated a pandemic with alarming rates of fatality worldwide. This situation has had a major impact on clinical laboratories that have attempted to answer the urgent need for diagnostic tools, since the identification of coronavirus disease 2019 (COVID-19). Development of a reliable serological diagnostic immunoassay, with high levels of sensitivity and specificity to detect SARS-CoV-2 antibodies with improved differential diagnosis from other circulating viruses, is mandatory.
METHODS
METHODS
An enzyme-linked immunosorbent assay (ELISA) using whole inactivated virus cultured in vitro, was developed to detect viral antigens. WB and ELISA investigations were carried out with sera of convalescent patients and negative sera samples. Both analyses were concurrently performed with recombinant MABs to verify the findings.
RESULTS
RESULTS
Preliminary data from 10 sera (5 patients with COVID-19, and 5 healthy controls) using this immunoassay are very promising, successfully identifying all of the confirmed SARS-CoV-2-positive individuals.
CONCLUSION
CONCLUSIONS
This ELISA appears to be a specific and reliable method for detecting COVID-19 antibodies (IgG, IgM, and IgA), and a useful tool for identifying individuals which have developed immunity to the virus.
Identifiants
pubmed: 33608968
doi: 10.1002/jcla.23735
pmc: PMC7995104
doi:
Substances chimiques
Antibodies, Viral
0
Antigens, Viral
0
Coronavirus Nucleocapsid Proteins
0
Phosphoproteins
0
Spike Glycoprotein, Coronavirus
0
nucleocapsid phosphoprotein, SARS-CoV-2
0
spike protein, SARS-CoV-2
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e23735Informations de copyright
© 2021 The Authors. Journal of Clinical Laboratory Analysis published by Wiley Periodicals LLC.
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