Jpn. J. Infect. Dis., 61 (3), 196-201, 2008

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Original Article

Analysis of Monthly Isolation of Respiratory Viruses from Children by Cell Culture Using a Microplate Method: a Two-Year Study from 2004 to 2005 in Yamagata, Japan

Katsumi Mizuta*, Chieko Abiko, Yoko Aoki, Asuka Suto, Hitoshi Hoshina, Tsutomu Itagaki1, Noriko Katsushima2, Yoko Matsuzaki3, Seiji Hongo3, Masahiro Noda4, Hirokazu Kimura5 and Katsumi Ootani

Department of Microbiology, Yamagata Prefectural Institute of Public Health, Yamagata 990-0031; 1Yamanobe Pediatric Clinic, Yamagata 990-0301; 2Katsushima Pediatric Clinic, Yamagata 990-2461; 3Department of Infectious Diseases, Yamagata University Faculty of Medicine, Yamagata 990-9585; and 4Department of Virology III and 5Infectious Diseases Surveillance Center, National Institute of Infectious Diseases, Tokyo 208-0011, Japan

(Received October 15, 2007. Accepted March 17, 2008)

*Corresponding author: Mailing address: Department of Microbiology, Yamagata Prefectural Institute of Public Health, Tokamachi 1-6-6, Yamagata, Yamagata 990-0031, Japan. Tel: +81-23-627-1109, Fax: +81-23-641-7486, E-mail:

SUMMARY: Although well over 200 viral agents have been implicated in acute respiratory infections (ARIs) among children, no system able to detect such a wide range of viruses has been established. Between January 2004 and December 2005, a modified microplate method, including HEF, HEp-2, Vero E6, MDCK, RD-18S, and GMK cell lines (HHVe6MRG plate), was adopted to isolate viruses. A total of 1,551 viruses were isolated, representing both outbreaks and sporadic cases, from 4,107 nasopharyngeal specimens, at monthly isolation rates of 22.3 to 52.6%. Influenza, parainfluenza, respiratory syncytial (RS), and mumps viruses, and human metapneumovirus, enterovirus, parechovirus, rhinovirus, adenovirus, herpesvirus, and cytomegalovirus were all isolated. The use of multiple cell lines increased the isolation rates of most of these viruses. The findings showed that ARIs due to a number of respiratory viruses occurred across all seasons in succession and/or concurrently in children in the community. These data will help clinicians determine in which seasons and for which age groups they should use the rapid diagnostic test kits available for influenza virus, RS virus, and adenovirus. In conclusion, we verified that the modified microplate method was able to clarify the etiology and epidemiology of numerous viruses isolated from children with ARI.

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