A competitive ELISA for species-independent detection of Crimean-Congo hemorrhagic fever virus specific antibodies.
Crimean-Congo hemorrhagic fever virus (CCHFV) circulates in many countries of Asia, Africa, and Europe. CCHFV can cause a severe hemorrhagic fever in humans with case-fatality rates of up to 80%. CCHF is considered to be one of the major emerging diseases spreading to and within Europe. Ticks of the genus Hyalomma function as vector as well as natural reservoir of CCHFV. Ticks feed on various domestic animals (e.g. cattle, sheep, goats) and on wildlife (e.g. hares, hedgehogs). Those animal species play an important role in the life cycle of the ticks as well as in amplification of CCHFV. Here we present a competitive ELISA (cELISA) for the species-independent detection of CCHFV-specific antibodies. For this purpose nucleocapsid (N) protein specific monoclonal antibodies (mAbs) were generated against an Escherichia coli (E. coli) expressed CCHFV N-protein. Thirty-three mAbs reacted with homologous and heterologous recombinant CCHFV antigens in ELISA and Western blot test and 20 of those 33 mAbs reacted additionally in an immunofluorescence assay with eukaryotic cells expressing the N-protein. Ten mAbs were further characterized in a prototype of the cELISA and nine of them competed with positive control sera of bovine origin. The cELISA was established by using the mAb with the strongest competition. For the validation, 833 sera from 12 animal species and from humans were used. The diagnostic sensitivity and specificity of the cELISA was determined to be 95% and 99%, respectively, and 2% of the sera gave inconclusive results. This cELISA offers the possibility for future large-scale screening approaches in various animal species to evaluate their susceptibility to CCHFV infection and to identify and monitor the occurrence of CCHFV.
Institute of Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, Greifswald-Insel Riems, Germany.
Isolde
I
Schuster
Institute of Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, Greifswald-Insel Riems, Germany.
Marc
M
Mertens
Institute of Immunology, Friedrich-Loeffler-Institut, Greifswald-Insel Riems, Germany.
Bernd
B
Köllner
Institute of Immunology, Friedrich-Loeffler-Institut, Greifswald-Insel Riems, Germany.
Tomáš
T
Korytář
Institute of Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, Greifswald-Insel Riems, Germany.
Markus
M
Keller
Department of Experimental Animal Facilities and Biorisk Management, Friedrich-Loeffler-Institut, Greifswald-Insel Riems, Germany.
Bärbel
B
Hammerschmidt
Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, Greifswald-Insel Riems, Germany.
Thomas
T
Müller
Unit Antiviral Strategies Antivirales, WHO Collaborative Centre for Viral Haemorrhagic Fevers and Arboviruses, OIE Reference Laboratory for RVFV and CCHFV, Institut Pasteur, Paris, France.
Noël
N
Tordo
Unit Virology; ANSES-Laboratoire de Lyon, Lyon, France.
Philippe
P
Marianneau
Institute of Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, Greifswald-Insel Riems, Germany.
Claudia
C
Mroz
Institute of Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, Greifswald-Insel Riems, Germany.
Melanie
M
Rissmann
Institute of Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, Greifswald-Insel Riems, Germany.
Eileen
E
Stroh
Institute of Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, Greifswald-Insel Riems, Germany.
Lisa
L
Dähnert
Chair of Food Safety, Faculty of Veterinary Medicine, Ludwig-Maximilians-University Munich (LMU), Oberschleissheim, Germany.
Felicitas
F
Hammerschmidt
Institute of Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, Greifswald-Insel Riems, Germany.
Rainer G
RG
Ulrich
Institute of Novel and Emerging Infectious Diseases, Friedrich-Loeffler-Institut, Greifswald-Insel Riems, Germany. Electronic address: martin.groschup@fli.de.
Martin H
MH
Groschup
D000349:Africa / Q000453:epidemiology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology; D001208:Asia / Q000453:epidemiology; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics; D005060:Europe / Q000453:epidemiology; D005455:Fluorescent Antibody Technique; D006041:Goats; D003225:Hemorrhagic Fever Virus, Crimean-Congo / Q000235:genetics / Q000276:immunology*; D006479:Hemorrhagic Fever, Crimean / Q000175:diagnosis* / Q000453:epidemiology / Q000276:immunology / Q000662:veterinary; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000276:immunology*; D012367:RNA, Viral; D012680:Sensitivity and Specificity; D012756:Sheep; D013987:Ticks / Q000821:virology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D007074:Immunoglobulin G; D019590:Nucleocapsid Proteins; D012367:RNA, Viral
Amplifying hosts; CCHFV; Competitive ELISA; Crimean-Congo hemorrhagic fever; Monoclonal antibody; Nucleocapsid protein