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Journal of Clinical Microbiology, February 1998, p. 340-344, Vol. 36, No. 2
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Application of Subtype-Specific Monoclonal Antibodies for Rapid Detection and Identification of Influenza A and B Viruses

Minoru Ueda,1 Akiko Maeda,2 Naoko Nakagawa,3 Tetsuo Kase,2 Ritsuko Kubota,2 Hikaru Takakura,1 Atsushi Ohshima,1 and Yoshinobu Okuno2,*

Biotechnology Research Laboratories, Takara Shuzo Co., Ltd., 3-4-1 Seta, Otsu, Shiga,1 and Division of Virology2 and Division of Bacteriology,3 Department of Public Health, Osaka Prefectural Institute of Public Health, 3-69, 1-Chome, Nakamichi, Higashinari-ku, Osaka 537 Japan

Received 14 July 1997/Returned for modification 10 October 1997/Accepted 5 November 1997

We established a rapid method for the identification of influenza A and B virus strains: the peroxidase-antiperoxidase (PAP) staining method with two subtype-specific murine monoclonal antibodies, C179 (H1 and H2 specific) and F49 (H3 specific), and an anti-influenza B virus rabbit polyclonal serum. The types and subtypes of 160 strains were examined, and 158 strains were identified to be the same by the hemagglutination-inhibition (HI) test and the PAP method. In contrast to the results by the HI test, two strains were revealed to be a mixture of two subtypes (H1 and H3) by the PAP method, which was confirmed by plaque cloning. We further analyzed clinical specimens by the PAP method by directly inoculating specimens into Madin-Darby canine kidney cells in microplates. After 40 h of incubation, the types and subtypes of viruses in 52 of 152 specimens were clearly identified. Since the reactivities of the two monoclonal antibodies are not influenced by the antigenic drift of influenza virus, the newly developed method should be applicable not only for rapid diagnosis but also for the epidemiological study of influenza.

* Corresponding author. Mailing address: Division of Virology, Department of Public Health, Osaka Prefectural Institute of Public Health, 3-69, 1-chome, Nakamichi, Higashinari-ku, Osaka 537, Japan. Phone: 81-6-972-1321. Fax: 81-6-972-2393. E-mail: okuno{at}iph.pref.osaka.jp.

Journal of Clinical Microbiology, February 1998, p. 340-344, Vol. 36, No. 2
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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