tetano
Editor, Senior Moderator
Eur J Clin Microbiol Infect Dis
. 2021 Mar 3.
doi: 10.1007/s10096-021-04203-8. Online ahead of print.
Automated Western immunoblotting detection of anti-SARS-CoV-2 serum antibodies
Sophie Edouard[SUP] 1 2 [/SUP], Rita Jaafar[SUP] 1 2 [/SUP], Nicolas Orain[SUP] 1 [/SUP], Philippe Parola[SUP] 1 3 [/SUP], Philippe Colson[SUP] 1 2 [/SUP], Bernard La Scola[SUP] 1 2 [/SUP], Pierre-Edouard Fournier[SUP] 1 3 [/SUP], Didier Raoult[SUP] 1 2 [/SUP], Michel Drancourt[SUP] 4 5 [/SUP]
Affiliations
Abstract
ELISA and chemiluminescence serological assays for COVID-19 are currently incorporating only one or two SARS-CoV-2 antigens. We developed an automated Western immunoblotting as a complementary serologic assay for COVID-19. The Jess[SUP]TM[/SUP] Simple Western system, an automated capillary-based assay, was used, incorporating an inactivated SARS-CoV-2 lineage 20a strain as the source of antigen, and total immunoglobulins (IgG, IgM, IgA) detection. In total, 602 sera were tested including 223 from RT-PCR-confirmed COVID-19 patients, 76 from patients diagnosed with seasonal HCoVs and 303 from coronavirus-negative control sera. We also compared this assay with the EUROIMMUN? SARS-CoV-2 IgG ELISA kit. Among 223 sera obtained from RT-PCR-confirmed COVID-19 patients, 180/223 (81%) exhibited reactivity against the nucleocapsid and 70/223 (31%) against the spike protein. Nucleocapsid reactivity was further detected in 9/76 (14%) samples collected from patients diagnosed with seasonal HCoVs and in 15/303 (5%) coronavirus-negative control samples. In the subset of sera collected more than 2 weeks after the onset of symptoms, the sensitivity was 94% and the specificity 93%, the latter value probably reflecting cross-reactivity of SARS-CoV-2 with other coronaviruses. The automated Western immunoblotting presented a substantial agreement (90%) with the compared ELISA (Cohen's Kappa=0.64). Automated Western immunoblotting may be used as a second line test to monitor exposure of people to HCoVs including SARS-CoV-2.
Keywords: Automated Western immunoblotting; COVID-19; Cross-reactivity; SARS-CoV-2; Serology.
. 2021 Mar 3.
doi: 10.1007/s10096-021-04203-8. Online ahead of print.
Automated Western immunoblotting detection of anti-SARS-CoV-2 serum antibodies
Sophie Edouard[SUP] 1 2 [/SUP], Rita Jaafar[SUP] 1 2 [/SUP], Nicolas Orain[SUP] 1 [/SUP], Philippe Parola[SUP] 1 3 [/SUP], Philippe Colson[SUP] 1 2 [/SUP], Bernard La Scola[SUP] 1 2 [/SUP], Pierre-Edouard Fournier[SUP] 1 3 [/SUP], Didier Raoult[SUP] 1 2 [/SUP], Michel Drancourt[SUP] 4 5 [/SUP]
Affiliations
- PMID: 33660134
- DOI: 10.1007/s10096-021-04203-8
Abstract
ELISA and chemiluminescence serological assays for COVID-19 are currently incorporating only one or two SARS-CoV-2 antigens. We developed an automated Western immunoblotting as a complementary serologic assay for COVID-19. The Jess[SUP]TM[/SUP] Simple Western system, an automated capillary-based assay, was used, incorporating an inactivated SARS-CoV-2 lineage 20a strain as the source of antigen, and total immunoglobulins (IgG, IgM, IgA) detection. In total, 602 sera were tested including 223 from RT-PCR-confirmed COVID-19 patients, 76 from patients diagnosed with seasonal HCoVs and 303 from coronavirus-negative control sera. We also compared this assay with the EUROIMMUN? SARS-CoV-2 IgG ELISA kit. Among 223 sera obtained from RT-PCR-confirmed COVID-19 patients, 180/223 (81%) exhibited reactivity against the nucleocapsid and 70/223 (31%) against the spike protein. Nucleocapsid reactivity was further detected in 9/76 (14%) samples collected from patients diagnosed with seasonal HCoVs and in 15/303 (5%) coronavirus-negative control samples. In the subset of sera collected more than 2 weeks after the onset of symptoms, the sensitivity was 94% and the specificity 93%, the latter value probably reflecting cross-reactivity of SARS-CoV-2 with other coronaviruses. The automated Western immunoblotting presented a substantial agreement (90%) with the compared ELISA (Cohen's Kappa=0.64). Automated Western immunoblotting may be used as a second line test to monitor exposure of people to HCoVs including SARS-CoV-2.
Keywords: Automated Western immunoblotting; COVID-19; Cross-reactivity; SARS-CoV-2; Serology.