title: Development of an antibody capture ELISA using inactivated Ebola Zaire Makona virus.
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.
authors.affiliation: Institute of Virology, Philipps University Marburg, Hans-Meerwein-Str 2, 35043, Marburg, Germany.|German Center for Infection Research (DZIF), Partner Site Giessen-Marburg-Langen, Hans-Meerwein-Str 2, 35043, Marburg, Germany.
authors.forename: Verena
authors.initials: V
authors.lastname: Krähling
authors.affiliation: Institute of Virology, Philipps University Marburg, Hans-Meerwein-Str 2, 35043, Marburg, Germany.|German Center for Infection Research (DZIF), Partner Site Giessen-Marburg-Langen, Hans-Meerwein-Str 2, 35043, Marburg, Germany.
authors.forename: Dirk
authors.initials: D
authors.lastname: Becker
authors.affiliation: Institute of Virology, Philipps University Marburg, Hans-Meerwein-Str 2, 35043, Marburg, Germany.
authors.forename: Cornelius
authors.initials: C
authors.lastname: Rohde
authors.affiliation: Institute of Virology, Philipps University Marburg, Hans-Meerwein-Str 2, 35043, Marburg, Germany.|German Center for Infection Research (DZIF), Partner Site Giessen-Marburg-Langen, Hans-Meerwein-Str 2, 35043, Marburg, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Eickmann
authors.affiliation: Institute of Virology, Philipps University Marburg, Hans-Meerwein-Str 2, 35043, Marburg, Germany.|German Center for Infection Research (DZIF), Partner Site Giessen-Marburg-Langen, Hans-Meerwein-Str 2, 35043, Marburg, Germany.
authors.forename: Yonca
authors.initials: Y
authors.lastname: Eroğlu
authors.affiliation: Institute of Virology, Philipps University Marburg, Hans-Meerwein-Str 2, 35043, Marburg, Germany.
authors.forename: Astrid
authors.initials: A
authors.lastname: Herwig
authors.affiliation: Bernhard Nocht Institute for Tropical Medicine, Bernhard-Nocht-Str. 74, 20359, Hamburg, Germany.
authors.forename: Romy
authors.initials: R
authors.lastname: Kerber
authors.affiliation: Institute of Virology, Philipps University Marburg, Hans-Meerwein-Str 2, 35043, Marburg, Germany.
authors.forename: Katharina
authors.initials: K
authors.lastname: Kowalski
authors.affiliation: Institute of Virology, Philipps University Marburg, Hans-Meerwein-Str 2, 35043, Marburg, Germany.|German Center for Infection Research (DZIF), Partner Site Giessen-Marburg-Langen, Hans-Meerwein-Str 2, 35043, Marburg, Germany.
authors.forename: Júlia
authors.initials: J
authors.lastname: Vergara-Alert
authors.affiliation: Institute of Virology, Philipps University Marburg, Hans-Meerwein-Str 2, 35043, Marburg, Germany. becker@staff.uni-marburg.de.|German Center for Infection Research (DZIF), Partner Site Giessen-Marburg-Langen, Hans-Meerwein-Str 2, 35043, Marburg, Germany. becker@staff.uni-marburg.de.
authors.forename: Stephan
authors.initials: S
authors.lastname: Becker
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology*; D002460:Cell Line; D029043:Ebolavirus / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000592:standards; D019142:Hemorrhagic Fever, Ebola / Q000175:diagnosis* / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D000914:Antibodies, Viral; D007074:Immunoglobulin G
keywords: Clinical diagnostics; ELISA; Ebola virus; Ebola virus vaccination; Seroconversion