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Applied and Environmental Microbiology, October 2003, p. 5782-5786, Vol. 69, No. 10
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.10.5782-5786.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Detection, Quantitation, and Phylogenetic Analysis of Noroviruses in Japanese Oysters

Tomoko Nishida,1 Hirokazu Kimura,2* Mika Saitoh,2 Michiyo Shinohara,3 Masahiko Kato,4 Shinji Fukuda,5 Tetsuya Munemura,6 Toshiyuki Mikami,7 Ayumi Kawamoto,8 Miho Akiyama,9 Yumiko Kato,9 Kanako Nishi,10 Kunihisa Kozawa,2 and Osamu Nishio9

Yamaguchi Prefectural Research Institute of Public Health, Yamaguchi,1 Gunma Prefectural Institute of Public Health and Environmental Sciences and Gunma University School of Medicine, Gunma,2 Saitama Prefectural Institute of Public Health, Saitama,3 Department of Pediatrics,4 Hiroshima Prefectural Institute of Public Health and Environment, Hiroshima,5 Yokohama City Institute of Public Health, Kanagawa,6 Aomori Prefectural Institute of Public Health and Environment, Aomori,7 Tottori Prefectural Institute of Public Health and Environmental Science, Tottori,8 National Institute of Public Health, Tokyo,9 Mie Prefectural Institute of Public Health, Mie, Japan,10

Received 13 January 2003/ Accepted 9 July 2003

Noroviruses (NVs) cause many cases of oyster- or clam-associated gastroenteritis in various countries. We collected 191 samples from Japanese oysters intended for raw consumption that had been harvested from the sea in two different areas between December 2001 and February 2002. To detect, quantitate, and phylogenetically analyze the NV genome in purified concentrates from the stomachs and digestive diverticula of these oysters, we amplified the NV capsid gene by reverse transcription-PCR. Phylogenetic analysis was performed by using the neighbor-joining method. We detected the NV genome in 17 of 191 oysters (9%). Phylogenetic analysis indicated genogroup I (Norwalk virus type) in 3 of the 17 oysters and genogroup II (Snow Mountain virus type) in the other 14. Both genogroups showed wide genetic diversity. To quantify the NV capsid gene in these oysters, we performed real-time PCR using genogroup-specific probes. More than 102 copies of the NV genome were detected in 11 of 17 oysters. The results suggested that about 10% of Japanese oysters intended for raw consumption harbored NVs, and more than 50% of those oysters in which NVs were detected had a large amount.

* Corresponding author. Mailing address: Gunma Prefectural Institute of Public Health and Environmental Sciences, 378 Kamioki, Maebashi, Gunma 371-0052, Japan. Phone: 81-27-232-4881. Fax: 81-27-234-8438. E-mail: kimura-hi{at}pref.gunma.jp.

Applied and Environmental Microbiology, October 2003, p. 5782-5786, Vol. 69, No. 10
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.10.5782-5786.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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