Development of a multiplex microsphere immunoassay for the detection of antibodies against highly pathogenic viruses in human and animal serum samples

Rebecca Surtees, Daniel Stern, Katharina Ahrens, Nicole Kromarek, Angelika Lander, Petra Kreher, Sabrina Weiss, Roger Hewson, Emma K. Punch, John N. Barr, Peter T. Witkowski, Emmanuel Couacy-Hymann, Andrea Marzi, Brigitte G. Dorner, Andreas Kurth*

*Corresponding author for this work

    Research output: Contribution to journalArticlepeer-review


    Surveillance of highly pathogenic viruses circulating in both human and animal populations is crucial to unveil endemic infections and potential zoonotic reservoirs. Monitoring the burden of disease by serological assay could be used as an early warning system for imminent outbreaks as an increased seroprevalance often precedes larger outbreaks. However, the multitude of highly pathogenic viruses necessitates the need to identify specific antibodies against several targets from both humans as well as from potential reservoir animals such as bats. In order to address this, we have developed a broadly reactive multiplex microsphere immunoassay (MMIA) for the detection of antibodies against several highly pathogenic viruses from both humans and animals. To this aim, nucleoproteins (NP) of Ebola virus (EBOV), Marburg virus (MARV) and nucleocapsid proteins (NP) of Crimean-Congo haemorrhagic fever virus, Rift Valley fever virus and Dobrava-Belgrade hantavirus were employed in a 5-plex assay for IgG detection. After optimisation, specific binding to each respective NP was shown by testing sera from humans and non-human primates with known infection status. The usefulness of our assay for serosurveillance was shown by determining the immune response against the NP antigens in a panel of 129 human serum samples collected in Guinea between 2011 and 2012 in comparison to a panel of 88 sera from the German blood bank. We found good agreement between our MMIA and commercial or in-house reference methods by ELISA or IIFT with statistically significant higher binding to both EBOV NP and MARV NP coupled microspheres in the Guinea panel. Finally, the MMIA was successfully adapted to detect antibodies from bats that had been inoculated with EBOV- and MARV- virus-like particles, highlighting the versatility of this technique and potentially enabling the monitoring of wildlife as well as human populations with this assay. We were thus able to develop and validate a sensitive and broadly reactive high-throughput serological assay which could be used as a screening tool to detect antibodies against several highly pathogenic viruses.

    Original languageEnglish
    Article numbere0008699
    Pages (from-to)1-23
    Number of pages23
    JournalPLoS Neglected Tropical Diseases
    Issue number10
    Publication statusPublished - Oct 2020

    Bibliographical note

    Funding Information:
    This research was funded by the Robert Koch Institute, DFG grant KR 1293/9-1 (SW, PTW, sampling sera in Guinea) and also in part supported by the Intramural Research Program, NIAID, NIH, USA (AM). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

    Publisher Copyright:
    © 2020, Public Library of Science. All rights reserved.


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