tetano
Editor, Senior Moderator
Pathogens
. 2020 Sep 28;9(10):E803.
doi: 10.3390/pathogens9100803.
Development and Optimization of In-house ELISA for Detection of Human IgG Antibody to SARS-CoV-2 Full Length Spike Protein
Thamir A Alandijany[SUP] 1 2 [/SUP], Sherif A El-Kafrawy[SUP] 1 2 [/SUP], Ahmed M Tolah[SUP] 1 [/SUP], Sayed S Sohrab[SUP] 1 2 [/SUP], Arwa A Faizo[SUP] 1 2 [/SUP], Ahmed M Hassan[SUP] 1 [/SUP], Tagreed L Alsubhi[SUP] 1 [/SUP], Norah A Othman[SUP] 1 [/SUP], Esam I Azhar[SUP] 1 2 [/SUP]
Affiliations
Abstract
The ongoing coronavirus disease 19 (COVID-19) pandemic, caused by the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), poses a threat to human health. Despite this, many affected countries are now in the process of gradual lifting of COVID-19 restrictions that were initially implemented in response to the pandemic. The success of the so-called "exit strategy" requires continued surveillance of virus circulation in the community and evaluation of the prevalence of protective immunity among population. Serology tests are valuable tools for these purposes. Herein, SARS-CoV-2 full-length spike (S) recombinant protein was utilized to develop and optimize an indirect enzyme-linked immunoassay (ELISA) that enables a reliable detection of virus-specific IgG antibody in human sera. Importantly, the performance of this assay was evaluated utilizing micro-neutralization (MN) assay as a reference test. Our developed ELISA offers 100% sensitivity, 98.4% specificity, 98.8% agreement, and high overall accuracy. Moreover, the optical density (OD) values of positive samples significantly correlated with their MN titers. The assay specifically detects human IgG antibodies directed against SARS-CoV-2, but not those to Middle East respiratory syndrome coronavirus (MERS-CoV) or human coronavirus HKU1 (HCoV-HKU1). The availability of this in-house ELISA protocol would be valuable for various diagnostic and epidemiological applications.
Keywords: COVID-19; ELISA; SARS-CoV-2; immunoassay.
. 2020 Sep 28;9(10):E803.
doi: 10.3390/pathogens9100803.
Development and Optimization of In-house ELISA for Detection of Human IgG Antibody to SARS-CoV-2 Full Length Spike Protein
Thamir A Alandijany[SUP] 1 2 [/SUP], Sherif A El-Kafrawy[SUP] 1 2 [/SUP], Ahmed M Tolah[SUP] 1 [/SUP], Sayed S Sohrab[SUP] 1 2 [/SUP], Arwa A Faizo[SUP] 1 2 [/SUP], Ahmed M Hassan[SUP] 1 [/SUP], Tagreed L Alsubhi[SUP] 1 [/SUP], Norah A Othman[SUP] 1 [/SUP], Esam I Azhar[SUP] 1 2 [/SUP]
Affiliations
- PMID: 32998438
- DOI: 10.3390/pathogens9100803
Abstract
The ongoing coronavirus disease 19 (COVID-19) pandemic, caused by the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), poses a threat to human health. Despite this, many affected countries are now in the process of gradual lifting of COVID-19 restrictions that were initially implemented in response to the pandemic. The success of the so-called "exit strategy" requires continued surveillance of virus circulation in the community and evaluation of the prevalence of protective immunity among population. Serology tests are valuable tools for these purposes. Herein, SARS-CoV-2 full-length spike (S) recombinant protein was utilized to develop and optimize an indirect enzyme-linked immunoassay (ELISA) that enables a reliable detection of virus-specific IgG antibody in human sera. Importantly, the performance of this assay was evaluated utilizing micro-neutralization (MN) assay as a reference test. Our developed ELISA offers 100% sensitivity, 98.4% specificity, 98.8% agreement, and high overall accuracy. Moreover, the optical density (OD) values of positive samples significantly correlated with their MN titers. The assay specifically detects human IgG antibodies directed against SARS-CoV-2, but not those to Middle East respiratory syndrome coronavirus (MERS-CoV) or human coronavirus HKU1 (HCoV-HKU1). The availability of this in-house ELISA protocol would be valuable for various diagnostic and epidemiological applications.
Keywords: COVID-19; ELISA; SARS-CoV-2; immunoassay.