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Clinical and Diagnostic Laboratory Immunology, January 2005, p. 202-205, Vol. 12, No. 1
1071-412X/05/$08.00+0     doi:10.1128/CDLI.12.1.202-205.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Immunofluorescence Assay for Detection of Human Metapneumovirus-Specific Antibodies by Use of Baculovirus-Expressed Fusion Protein

Nobuhisa Ishiguro,1 Takashi Ebihara,1 Rika Endo,1 Xiaoming Ma,1 Ryo Shirotsuki,1 Susumu Ochiai,2 Hiroaki Ishiko,2 and Hideaki Kikuta1*

Department of Pediatrics, Hokkaido University Graduate School of Medicine, Sapporo,1 Department of Infectious Diseases, Mitsubishi Kagaku Bio-Clinical Laboratories, Tokyo, Japan2

Received 25 July 2004/ Returned for modification 27 September 2004/ Accepted 13 October 2004

Human metapneumovirus (hMPV) has recently been identified as an etiological agent of acute respiratory infections. The hMPV fusion (F) protein has been indicated to be a major antigenic determinant that mediates effective neutralization and protection against hMPV infection. We developed a new immunofluorescence assay (IFA) using Trichoplusia ni (Tn5) insect cells infected with a recombinant baculovirus-expressing hMPV F protein (Bac-F IFA). A total of 200 serum samples from Japanese people 1 month to 41 years old were tested for immunoglobulin G antibodies to hMPV F protein by Bac-F IFA. The results were compared with those of the conventional IFA based on hMPV-infected LLC-MK2 cells (hMPV IFA). The titers obtained by the two IFAs correlated well (correlation coefficient of 0.88), and the concordance of seroreactivities between the two IFAs was 91% ({kappa} = 0.76). For 192 of the 200 serum samples, the titers obtained by the Bac-F IFA were equal to or higher than those obtained by the hMPV IFA. These results indicated that the Bac-F IFA was more sensitive than the hMPV IFA and that the majority of the antibodies detected by the hMPV IFA reacted with the hMPV F protein. The Bac-F IFA is a more reliable, sensitive, and specific method for the detection of hMPV antibodies than is the hMPV IFA.

* Corresponding author. Mailing address: Department of Pediatrics, Hokkaido University Graduate School of Medicine, N-15, W-7, Kita-ku, Sapporo 060-8638, Japan. Phone: 81-11-706-5954. Fax: 81-11-706-7898. E-mail: hide-ki{at}med.hokudai.ac.jp.

Clinical and Diagnostic Laboratory Immunology, January 2005, p. 202-205, Vol. 12, No. 1
1071-412X/05/$08.00+0     doi:10.1128/CDLI.12.1.202-205.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.





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