Development of an antibody capture ELISA using inactivated Ebola Zaire Makona virus


The 2014 Zaire Ebola virus (ZEBOV) outbreak in West Africa represents an international public health concern. Highly sensitive and precise diagnostic tools are needed. In the present study, we developed a ZEBOV-specific enzyme-linked immunosorbent assay (ELISA) using inactivated ZEBOV isolate Makona from March 2014. Mock antigen was used to address nonspecific binding. Specificity, reproducibility and precision were determined to measure assay performance. The ZEBOV ELISA proved to be specific (96 %), reproducible and precise (Intra-assay CV 8 %, Inter-assay CV 18 %). Using the human monoclonal antibody KZ52, we showed that the ELISA was able to detect conformation-specific antibodies. Monitoring antibody development in 29 PCR-positive EBOV disease (EVD) patients revealed seroconversion in all cases. In addition, the ELISA was used to detect ZEBOV glycoprotein (GP)-specific antibodies in a vaccinated volunteer from day 14 until 5 years post-vaccination with a VSV-ZEBOV candidate vaccine. The results demonstrate the high reproducibility, specificity and sensitivity of this newly developed ELISA, which is suitable for the detection of specific antibody responses directed against different ZEBOV proteins in EVD patients and against the ZEBOV surface glycoprotein GP in vaccinated individuals.

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This work was supported by the German Center for Infection Research (DZIF), the Wellcome Trust Foundation through the World Health Organization: “Immunogenicity of Ebola Virus vaccine” (Grant Number: SPHQ14-LOA-296), the Federal Ministry of Education and Research (EBOKON), the European Union: EVIDENT “Ebola Virus Disease—correlates of protection, determinants of outcome, and clinical management” (Horizon 2020 Grant Agreement No.: 666100 and service contract IFS/2011/272-372), as well as the Jürgen-Manchot-Foundation through a stipend to CR and the German Research Foundation (DFG, SPP1596). We especially thank Gotthard Ludwig and Michael Schmidt for their expert technical support with the BSL4 procedures at Philipps University of Marburg. We thank Carsten van Hammel, Ronny Kohlhoff, and Jörg Schmidt for the technical and administrative support. We also thank all of the voluntary donors for generously providing serum samples for scientific analyses as well as Petra Emmerich, Timo Wolf, and Marylyn M. Addo for providing some of the samples. We thank Stephanie Wurr und Elisa Pallasch for excellent assistance with biobanking in the BSL-4 laboratory at BNITM. The European Mobile Lab consortium: Miles W. Carroll, Roger Hewson, Joseph Akoi Bore, Raymond Koundouno, Saïd Abdellati, Babak Afrough, John Aiyepada, Patience Akhilomen, Danny Asogun, Barry Atkinson, Marlis Badusche, Amadou Bah, Simon Bate, Jan Baumann, Dirk Becker, Beate Becker-Ziaja, Anne Bocquin, Benny Borremans, Andrew Bosworth, Jan Peter Boettcher, Angela Cannas, Fabrizio Carletti, Concetta Castilletti, Simon Clark, Francesca Colavita, Sandra Diederich, Adomeh Donatus, Sophie Duraffour, Deborah Ehichioya, Heinz Ellerbrok, Maria Dolores Fenandez-Garcia, Alexandra Fizet, Erna Fleischmann, Sophie Gryseels, Antje Hermelink, Julia Hinzmann, Ute Hopf-Guevara, Yemisi Ighodalo, Lisa Jameson, Anne Kelterbaum, Zoltan Kis, Stefan Kloth, Claudia Kohl, Miša Korva, Annette Kraus, Eeva Kuisma, Andreas Kurth, Britta Liedigk, Christopher H. Logue, Anja Lüdtke, Piet Maes, James McCowen, Stéphane Mély, Marc Mertens, Silvia Meschi, Benjamin Meyer, Janine Michel, Peter Molkenthin, César Muñoz-Fontela, Doreen Muth, Edmund N. C. Newman, Didier Ngabo, Lisa Oestereich, Jennifer Okosun, Thomas Olokor, Racheal Omiunu, Emmanuel Omomoh, Elisa Pallasch, Bernadett Pályi, Jasmine Portmann, Thomas Pottage, Catherine Pratt, Simone Priesnitz, Serena Quartu, Julie Rappe, Johanna Repits, Martin Richter, Martin Rudolf, Andreas Sachse, Kristina Maria Schmidt, Gordian Schudt, Thomas Strecker, Ruth Thom, Stephen Thomas, Ekaete Tobin, Howard Tolley, Jochen Trautner, Tine Vermoesen, Inês Vitoriano, Matthias Wagner, Svenja Wolff, Constanze Yue, Maria Rosaria Capobianchi, Romy Kerber, Tatjana Avšič-Županc, Andreas Nitsche, Marc Strasser, Giuseppe Ippolito, Stephan Becker, Kilian Stoecker, Martin Gabriel, Hervé Raoul, Antonino Di Caro (executive board), Roman Wölfel (executive board) and Stephan Günther (executive board).

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Krähling, V., Becker, D., Rohde, C. et al. Development of an antibody capture ELISA using inactivated Ebola Zaire Makona virus. Med Microbiol Immunol 205, 173–183 (2016).

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  • Ebola virus
  • Clinical diagnostics
  • Ebola virus vaccination
  • Seroconversion