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Automated Western immunoblotting detection of anti-SARS-CoV-2 serum antibodies

Abstract : 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 TM 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.
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Submitted on : Monday, May 3, 2021 - 12:42:09 PM
Last modification on : Friday, April 1, 2022 - 3:55:48 AM

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Sophie Edouard, Rita Jaafar, Nicolas Orain, Philippe Parola, Philippe Colson, et al.. Automated Western immunoblotting detection of anti-SARS-CoV-2 serum antibodies. European Journal of Clinical Microbiology and Infectious Diseases, Springer Verlag, 2021, ⟨10.1007/s10096-021-04203-8⟩. ⟨hal-03215451⟩



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