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Journal of Clinical Microbiology, February 2000, p. 530-536, Vol. 38, No. 2
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Molecular Detection and Epidemiology of Sapporo-Like Viruses

Jan Vinjé,1 Hanneke Deijl,1 Reina van der Heide,1 David Lewis,2 Kjell-Olof Hedlund,3 Lennart Svensson,3 and Marion P. G. Koopmans1,*

National Institute of Public Health and the Environment, Bilthoven, The Netherlands1; Public Health Laboratory Services, Leeds, United Kingdom2; and Karolinska Institute, Solna, Sweden3

Received 6 July 1999/Returned for modification 21 September 1999/Accepted 5 November 1999

Sapporo-like viruses (SLVs) are associated with acute gastroenteritis in humans. Due to a limited supply of available reagents for diagnosis, little is known about the incidence and pathogenicity of these viruses. We have developed a first-generation generic reverse transcriptase (RT) PCR assay based on a single primer pair targeting the RNA polymerase gene. With this assay, 55 (93%) of the 59 stool specimens collected in a 10-year period of time (1988 to 1998) and containing typical caliciviruses by electron microscopy tested positive and could be confirmed by Southern hybridization. By phylogenetic analysis, most SLV strains could be classified into one of the three recently described genotypes. However, three samples clustered separately, forming a potential new genotype. We sequenced the complete capsid gene of one of the strains in this cluster: Hu/SLV/Stockholm/97/SE. Alignment of the capsid sequences showed 40 to 74% amino acid identity among strains of the different clusters. Phylogenetic analysis of the aligned sequences confirmed the placing of Hu/SLV/Stockholm/97/SE into a new distinct genetic cluster. This is the first report on the development of a broadly reactive RT-PCR assay for the detection of SLVs.

* Corresponding author. Mailing address: Research Laboratory for Infectious Diseases, Department of Virology, National Institute of Public Health and the Environment (R.I.V.M.), Antonie van Leeuwenhoeklaan 9, 3721 MA Bilthoven, The Netherlands. Phone: (31)-30-2743944. Fax: (31)-30-2744449. E-mail: Marion.Koopmans{at}rivm.nl.

Journal of Clinical Microbiology, February 2000, p. 530-536, Vol. 38, No. 2
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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