Jpn. J. Infect. Dis., 58, 88-94, 2005

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Original Article

Immunological Detection of Severe Acute Respiratory Syndrome Coronavirus by Monoclonal Antibodies

Kazuo Ohnishi, Masahiro Sakaguchi, Tomohiro Kaji, Kiyoko Akagawa, Tadayoshi Taniyama, Masataka Kasai, Yasuko Tsunetsugu-Yokota, Masamichi Oshima, Kiichi Yamamoto, Naomi Takasuka, Shu-ichi Hashimoto, Manabu Ato, Hideki Fujii, Yoshimasa Takahashi, Shigeru Morikawa1, Koji Ishii2, Tetsutaro Sata4, Hirotaka Takagi5, Shigeyuki Itamura3, Takato Odagiri3, Tatsuo Miyamura2, Ichiro Kurane1, Masato Tashiro3, Takeshi Kurata6, Hiroshi Yoshikura6 and Toshitada Takemori*

Department of Immunology, 1Department of Virology I, 2Department of Virology II, 3Department of Virology III, 4Department of Pathology and 5Division of Biosafety Control and Research, 6National Institute of Infectious Diseases, Tokyo 162-8640, Japan

(Received October 20, 2004. Accepted February 14, 2005)

*Corresponding author: Mailing address: Department of Immunology, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo 162-8640, Japan. Tel: +81-3-5285-1111, Fax: +81-5285-1150, E-mail: ttoshi@nih.go.jp


SUMMARY: In order to establish immunological detection methods for severe acute respiratory syndrome coronavirus (SARS-CoV), we established monoclonal antibodies directed against structural components of the virus. B cell hybridomas were generated from mice that were hyper-immunized with inactivated SARS-CoV virion. By screening 2,880 generated hybridomas, we established three hybridoma clones that secreted antibodies specific for nucleocapsid protein (N) and 27 clones that secreted antibodies specific for spike protein (S). Among these, four S-protein specific antibodies had in vitro neutralization activity against SARS-CoV infection. These monoclonal antibodies enabled the immunological detection of SARS-CoV by immunofluorescence staining, Western blot or immunohistology. Furthermore, a combination of monoclonal antibodies with different specificities allowed the establishment of a highly sensitive antigen-capture sandwich ELISA system. These monoclonal antibodies would be a useful tool for rapid and specific diagnosis of SARS and also for possible antibody-based treatment of the disease.

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