Journal
EUROPEAN JOURNAL OF CLINICAL MICROBIOLOGY & INFECTIOUS DISEASES
Volume 40, Issue 6, Pages 1309-1317Publisher
SPRINGER
DOI: 10.1007/s10096-021-04203-8
Keywords
SARS-CoV-2; COVID-19; Cross-reactivity; Serology; Automated Western immunoblotting
Categories
Funding
- IHU Mediterranee Infection, Marseille, France
- fondation Mediterranee Infection, Marseille, France
- ANR Investissements d'Avenir Mediterranee Infection [10-IAHU-03]
- [ANR-15-CE36-0004-01]
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ELISA and chemiluminescence serological assays for COVID-19 currently focus on one or two SARS-CoV-2 antigens, while an automated Western immunoblotting was developed as a complementary serologic assay, using an inactivated SARS-CoV-2 lineage 20a strain as the antigen source. The automated Western immunoblotting showed substantial agreement (90%) with the compared ELISA, with a sensitivity of 94% and a specificity of 93%, particularly in samples collected more than 2 weeks after symptom onset.
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 (R) 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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