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Journal of Clinical Microbiology, September 2001, p. 3267-3271, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3267-3271.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Detection of Ebola Viral Antigen by Enzyme-Linked Immunosorbent Assay Using a Novel Monoclonal Antibody to Nucleoprotein

Masahiro Niikura,1 Tetsuro Ikegami,2 Masayuki Saijo,1 Ichiro Kurane,1 Mary E. Miranda,3 and Shigeru Morikawa1,*

Department of Virology 1, National Institute of Infectious Diseases,1 and Department of Biomedical Science, Graduate School of Agricultural and Life Sciences, The University of Tokyo,2 Tokyo, Japan, and Research Institute for Tropical Medicine, Department of Health, Muntinlupa City, Philippines3

Received 13 November 2000/Returned for modification 2 March 2001/Accepted 19 June 2001

With the increase in international traffic, the risk of introducing rare but severe infectious diseases like Ebola hemorrhagic fever is increasing all over the world. However, the system for the diagnosis of Ebola virus infection is available in a limited number of countries. In the present study, we developed an Ebola virus antigen-detection enzyme-linked immunosorbent assay (ELISA) system using a novel monoclonal antibody (MAb) to the nucleoprotein (NP). This antibody recognized an epitope defined by a 26-amino-acid stretch near the C terminus of NP. In a sandwich ELISA system with the MAb, as little as 30 ng of purified recombinant NP (rNP) was detected. Although this MAb was prepared by immunization with rNP of subtype Zaire, it also reacted to the corresponding region of NP derived from the Reston and Sudan subtypes. These results suggest that our ELISA system should work with three of four Ebola subtypes. Furthermore, our ELISA system detected the NP in subtype Reston-infected monkey specimens, while the background level in noninfected specimens was very low, suggesting the usefulness of the ELISA for laboratory diagnosis with clinical specimens.

* Corresponding author. Mailing address: Special Pathogens Laboratory, Department of Virology 1, National Institute of Infectious Diseases, 4-7-1 Gakuen, Musashimurayama, 208-0011 Tokyo, Japan. Phone: 81-42-561-0771, ext. 791. Fax: 81-42-564-4881. E-mail: morikawa{at}nih.go.jp.

Journal of Clinical Microbiology, September 2001, p. 3267-3271, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3267-3271.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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Copyright © 2001 by the American Society for Microbiology. All rights reserved.