Format

Send to

Choose Destination
J Virol Methods. 2018 Jan;251:22-29. doi: 10.1016/j.jviromet.2017.10.008. Epub 2017 Oct 6.

Characterization of novel monoclonal antibodies against the MERS-coronavirus spike protein and their application in species-independent antibody detection by competitive ELISA.

Author information

1
Department of Virology I, National Institute of Infectious Diseases, Japan. Electronic address: fukushi@nih.go.jp.
2
Department of Virology I, National Institute of Infectious Diseases, Japan.
3
Department of Virology III, National Institute of Infectious Diseases, Japan.
4
Department of Pathology, National Institute of Infectious Diseases, Japan.
5
Department of Immunology, National Institute of Infectious Diseases, Japan.
6
Department of Biotechnology, College of Biological and Chemical Engineering, Addis Ababa Science and Technology University, Ethiopia.
7
Department of Veterinary Medicine, Nihon University, Japan.

Abstract

Since discovering the Middle East respiratory syndrome coronavirus (MERS-CoV) as a causative agent of severe respiratory illness in the Middle East in 2012, serological testing has been conducted to assess antibody responses in patients and to investigate the zoonotic reservoir of the virus. Although the virus neutralization test is the gold standard assay for MERS diagnosis and for investigating the zoonotic reservoir, it uses live virus and so must be performed in high containment laboratories. Competitive ELISA (cELISA), in which a labeled monoclonal antibody (MAb) competes with test serum antibodies for target epitopes, may be a suitable alternative because it detects antibodies in a species-independent manner. In this study, novel MAbs against the spike protein of MERS-CoV were produced and characterized. One of these MAbs was used to develop a cELISA. The cELISA detected MERS-CoV-specific antibodies in sera from MERS-CoV-infected rats and rabbits immunized with the spike protein of MERS-CoV. The MAb-based cELISA was validated using sera from Ethiopian dromedary camels. Relative to the neutralization test, the cELISA detected MERS-CoV-specific antibodies in 66 Ethiopian dromedary camels with a sensitivity and specificity of 98% and 100%, respectively. The cELISA and neutralization test results correlated well (Pearson's correlation coefficients=0.71-0.76, depending on the cELISA serum dilution). This cELISA may be useful for MERS epidemiological investigations on MERS-CoV infection.

KEYWORDS:

Competitive ELISA; Epidemiology; MERS coronavirus; Neutralizing antibody

PMID:
28993122
DOI:
10.1016/j.jviromet.2017.10.008
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center