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Journal of Clinical Microbiology, June 2007, p. 1912-1919, Vol. 45, No. 6
0095-1137/07/$08.00+0     doi:10.1128/JCM.01251-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Outbreak of Human Metapneumovirus Detected by Use of the Vero E6 Cell Line in Isolates Collected in Yamagata, Japan, in 2004 and 2005{triangledown}

C. Abiko,1 K. Mizuta,1* T. Itagaki,2 N. Katsushima,3 S. Ito,4 Y. Matsuzaki,5 M. Okamoto,6 H. Nishimura,6 Y. Aoki,1 T. Murata,1 H. Hoshina,1 S. Hongo,5 and K. Ootani1

Department of Microbiology, Yamagata Prefectural Institute of Public Health, Yamagata, Japan,1 Yamanobe Pediatric Clinic, Yamagata, Japan,2 Katsushima Pediatric Clinic, Yamagata, Japan,3 Department of Pediatrics, Shonai Hospital, Yamagata, Japan,4 Department of Infectious Diseases, Yamagata University School of Medicine, Yamagata, Japan,5 Virus Research Center, Clinical Research Division, Sendai Medical Center, Sendai, Japan6

Received 19 June 2006/ Returned for modification 24 September 2006/ Accepted 9 April 2007

A number of epidemiological studies have shown human metapneumovirus (hMPV) to be one of the most important viral agents associated with acute respiratory infections in humans. However, due to the difficulty in growing the virus, all epidemiological studies of hMPV infection have been performed on the basis of the molecular method. Thus, the development of a cell line suitable for the isolation of hMPV from clinical specimens is a crucial step for further research. Using the Vero E6 cell line, which could be stably maintained for 1 month without passage or medium change, we succeeded in isolating 79 strains from 4,112 specimens obtained in Yamagata, Japan, in 2004 and 2005. The total isolation rate was 1.9% (79/4,112). The monthly distribution revealed that hMPV infections occurred between February and April in 2004 and throughout most of the year in 2005. Phylogenetic analysis indicated that subgenogroup B2 was predominant in 2004, whereas three subgenogroups, A2, B1, and B2, had cocirculated in 2005. Although multiple subgenogroups cocirculated in 2005, each individual subgenogroup strain was found to predominate at specific sites. An infectivity assay of hMPV strains also indicated that the infection efficiency in Vero E6 cells was better than that in LLC-MK2 cells. Finally, we found that Vero E6 cells are useful for the isolation of hMPVs and that this utility might aid further research into hMPVs beyond the epidemiological data shown in this study.

* Corresponding author. Mailing address: Department of Microbiology, Yamagata Prefectural Institute of Public Health, Tokamachi, Yamagata 990-0031, Japan. Phone: 81-23-627-1109. Fax: 81-23-641-7486. E-mail: mizutak{at}pref.yamagata.jp

{triangledown} Published ahead of print on 25 April 2007.

Journal of Clinical Microbiology, June 2007, p. 1912-1919, Vol. 45, No. 6
0095-1137/07/$08.00+0     doi:10.1128/JCM.01251-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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