Development of indigenous IgG ELISA for the detection of anti-SARS-CoV-2 IgG.
Antibodies, Viral
/ blood
Betacoronavirus
/ immunology
COVID-19
COVID-19 Testing
Clinical Laboratory Techniques
/ methods
Coronavirus Infections
/ blood
Enzyme-Linked Immunosorbent Assay
Humans
Immunoglobulin G
/ blood
India
/ epidemiology
Pandemics
Pneumonia, Viral
/ blood
Predictive Value of Tests
Prevalence
ROC Curve
Reproducibility of Results
SARS-CoV-2
Seroepidemiologic Studies
COVID-19 - diagnosis - ELISA - human - IgG antibodies - microneutralization test - SARS-CoV-2 - standardization
Journal
The Indian journal of medical research
ISSN: 0971-5916
Titre abrégé: Indian J Med Res
Pays: India
ID NLM: 0374701
Informations de publication
Date de publication:
May 2020
May 2020
Historique:
entrez:
3
7
2020
pubmed:
3
7
2020
medline:
14
7
2020
Statut:
ppublish
Résumé
Since the beginning of the year 2020, the pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) impacted humankind adversely in almost all spheres of life. The virus belongs to the genus Betacoronavirus of the family Coronaviridae. SARS-CoV-2 causes the disease known as coronavirus disease 2019 (COVID-19) with mild-to-severe respiratory illness. The currently available diagnostic tools for the diagnosis of COVID-19 are mainly based on molecular assays. Real-time reverse transcription-polymerase chain reaction is the only diagnostic method currently recommended by the World Health Organization for COVID-19. With the rapid spread of SARS-CoV-2, it is necessary to utilize other tests, which would determine the burden of the disease as well as the spread of the outbreak. Considering the need for the development of such a screening test, an attempt was made to develop and evaluate an IgG-based ELISA for COVID-19. A total of 513 blood samples (131 positive, 382 negative for SARS-CoV-2) were collected and tested by microneutralization test (MNT). Antigen stock of SARS-CoV-2 was prepared by propagating the virus in Vero CCL-81 cells. An IgG capture ELISA was developed for serological detection of anti-SARS-CoV-2 IgG in serum samples. The end point cut-off values were determined by using receiver operating characteristic (ROC) curve. Inter-assay variability was determined. The developed ELISA was found to be 92.37 per cent sensitive, 97.9 per cent specific, robust and reproducible. The positive and negative predictive values were 94.44 and 98.14 per cent, respectively. This indigenously developed IgG ELISA was found to be sensitive and specific for the detection of anti-SARS-CoV-2 IgG in human serum samples. This assay may be used for determining seroprevalence of SARS-CoV-2 in a population exposed to the virus.
Sections du résumé
BACKGROUND & OBJECTIVES
OBJECTIVE
Since the beginning of the year 2020, the pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) impacted humankind adversely in almost all spheres of life. The virus belongs to the genus Betacoronavirus of the family Coronaviridae. SARS-CoV-2 causes the disease known as coronavirus disease 2019 (COVID-19) with mild-to-severe respiratory illness. The currently available diagnostic tools for the diagnosis of COVID-19 are mainly based on molecular assays. Real-time reverse transcription-polymerase chain reaction is the only diagnostic method currently recommended by the World Health Organization for COVID-19. With the rapid spread of SARS-CoV-2, it is necessary to utilize other tests, which would determine the burden of the disease as well as the spread of the outbreak. Considering the need for the development of such a screening test, an attempt was made to develop and evaluate an IgG-based ELISA for COVID-19.
METHODS
METHODS
A total of 513 blood samples (131 positive, 382 negative for SARS-CoV-2) were collected and tested by microneutralization test (MNT). Antigen stock of SARS-CoV-2 was prepared by propagating the virus in Vero CCL-81 cells. An IgG capture ELISA was developed for serological detection of anti-SARS-CoV-2 IgG in serum samples. The end point cut-off values were determined by using receiver operating characteristic (ROC) curve. Inter-assay variability was determined.
RESULTS
RESULTS
The developed ELISA was found to be 92.37 per cent sensitive, 97.9 per cent specific, robust and reproducible. The positive and negative predictive values were 94.44 and 98.14 per cent, respectively.
INTERPRETATION & CONCLUSIONS
CONCLUSIONS
This indigenously developed IgG ELISA was found to be sensitive and specific for the detection of anti-SARS-CoV-2 IgG in human serum samples. This assay may be used for determining seroprevalence of SARS-CoV-2 in a population exposed to the virus.
Identifiants
pubmed: 32611915
pii: IndianJMedRes_2020_151_5_444_286007
doi: 10.4103/ijmr.IJMR_2232_20
pmc: PMC7530443
doi:
Substances chimiques
Antibodies, Viral
0
Immunoglobulin G
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
444-449Déclaration de conflit d'intérêts
None
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