CVI Accepts, published online ahead of print on 24 June 2009

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Clin. Vaccine Immunol. doi:10.1128/CVI.00163-09
Copyright (c) 2009, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Characterization of Monoclonal Antibodies to Junin Virus Nucleocapsid protein and Application to the Diagnosis of Hemorrhagic Fever Caused by South American Arenaviruses

Mina Nakauchi, Shuetsu Fukushi, Masayuki Saijo, Tetsuya Mizutani, Agustín E. Ure, Victor Romanowski, Ichiro Kurane, and Shigeru Morikawa^*

Department of Virology 1, National Institute of Infectious Diseases, 4-7-1 Gakuen, Musashimurayama, Tokyo 208-0011, Japan; and Facultad de Ciencias Exactas, Instituto de Biotecnología y Biología Molecular, Universidad Nacional de La Plata, Consejo Nacional de Investigaciones Científicas y Técnicas (IBBM-UNLP-CONICET-CCT La Plata), La Plata, Argentina

^* To whom correspondence should be addressed. Email: morikawa{at}nih.go.jp.

The Junin virus (JUNV), Machupo virus, Guanarito virus, Sabia virus, and Chapare virus, are members of the Clade B New World arenaviruses and are the etiological agents of viral hemorrhagic fevers that occur in South America. In this study, we produced three monoclonal antibodies (MAbs) to recombinant nucleocapsid protein of JUNV, designated as MAbs C6-9, C11-12, and E4-2. The specificity of these MAbs was examined by enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay, and an epitope mapping method. Using these MAbs, we developed antigen (Ag)-capture ELISA systems. We showed that, by using MAb C6-9, JUNV antigen was specifically detected. On the other hand, by using MAb C11-12 or E-4-2, the antigens of all human pathogenic South American arenaviruses were detected. The combined use of these Ag-capture ELISA systems in the present study may be useful for the diagnosis of acute phase viral hemorrhagic fever due to infection by a South American arenavirus.