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

Verena Krähling, Dirk Becker, Cornelius Rohde, Markus Eickmann, Yonca Eroğlu, Astrid Herwig, Romy Kerber, Katharina Kowalski, Júlia Vergara-Alert, Stephan Becker, European Mobile Laboratory consortium, Angela Cannas, Fabrizio Carletti, Concetta Castilletti, Francesca Colavita, Silvia Meschi, Serena Quartu, Maria Rosaria Capobianchi, Giuseppe Ippolito, Antonino Di Caro

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish
Pages (from-to)173-83
Number of pages11
JournalMedical Microbiology and Immunology
Volume205
Issue number2
DOIs
Publication statusPublished - Apr 2016

Keywords

  • Animals
  • Antibodies, Viral
  • Cell Line
  • Ebolavirus
  • Enzyme-Linked Immunosorbent Assay
  • Hemorrhagic Fever, Ebola
  • Humans
  • Immunoglobulin G
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Journal Article
  • Research Support, Non-U.S. Gov't

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