The presence of antibodies against endemic coronaviruses has been
linked to disease severity after SARS-CoV-2 infection. Assays
capable of concomitantly detecting antibodies against endemic
coronaviridae such as OC43, 229E, NL63, and SARS-CoV-2 may help
to elucidate this question. We developed a serum screening
platform using a bead-based Western blot system called DigiWest,
capable of running hundreds of assays using microgram amounts of
protein prepared directly from different viruses.
Characterization of the immunoassay for detection of SARS-CoV-2
specific antibodies revealed a sensitivity of 90.3% and a
diagnostic specificity of 98.1%. Concordance analysis with the
SARS-CoV-2 immunoassays available by Roche, Siemens, and
Euroimmun indicates comparable assay performances
(Cohen’s κ ranging from 0.8874 to 0.9508).
Analogous assays for OC43, 229E, and NL63 were established and
combined into one multiplex with the SARS-CoV-2 assay.
Seroreactivity for different coronaviruses was detected with
high incidence, and the multiplex assay was adapted for serum
screening.
BACKGROUND: Seroreactivity against human endemic coronaviruses has been linked to disease severity after SARS-CoV-2 infection. Assays that are capable of concomitantly detecting antibodies against endemic coronaviridae such as OC43, 229E, NL63, and SARS-CoV-2 may help to elucidate this question. We set up a platform for serum-screening and developed a bead-based Western blot system, namely DigiWest, capable of running hundreds of assays using microgram amounts of protein prepared directly from different viruses.
METHODS: The parallelized and miniaturised DigiWest assay was adapted for detecting antibodies using whole protein extract prepared from isolated SARS-CoV-2 virus particles. After characterisation and optimization of the newly established test, whole virus lysates of OC43, 229E, and NL63 were integrated into the system.
RESULTS: The DigiWest-based immunoassay system for detection of SARS-CoV-2 specific antibodies shows a sensitivity of 87.2 % and diagnostic specificity of 100 %. Concordance analysis with the SARS-CoV-2 immunoassays available by Roche, Siemens, and Euroimmun indicates a comparable assay performance (Cohen's Kappa ranging from 0.8799-0.9429). In the multiplexed assay, antibodies against the endemic coronaviruses OC43, 229E, and NL63 were detected, displaying a high incidence of seroreactivity against these coronaviruses.
CONCLUSION: The DigiWest-based immunoassay, which uses authentic antigens from isolated virus particles, is capable of detecting individual serum responses against SARS-CoV-2 with high specificity and sensitivity in one multiplexed assay. It shows high concordance with other commercially available serologic assays. The DigiWest approach enables a concomitant detection of antibodies against different endemic coronaviruses and will help to elucidate the role of these possibly cross-reactive antibodies.
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