Molecular detection and epidemiology of Sapporo-like viruses.
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.
Vinjé
J
J
National Institute of Public Health and the Environment, Bilthoven, The Netherlands.
Deijl
H
H
van der Heide
R
R
Lewis
D
D
Hedlund
K O
KO
Svensson
L
L
Koopmans
M P
MP
D000328:Adult; D000368:Aged; D015139:Blotting, Southern; D002139:Caliciviridae / Q000737:chemistry / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D017250:Caliciviridae Infections / Q000453:epidemiology* / Q000821:virology; D002213:Capsid / Q000737:chemistry / Q000235:genetics; D002648:Child; D002675:Child, Preschool; D017931:DNA Primers; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D016208:Databases, Factual; D005243:Feces / Q000821:virology; D005759:Gastroenteritis / Q000453:epidemiology* / Q000821:virology*; D014644:Genetic Variation; D006801:Humans; D007223:Infant; D008854:Microscopy, Electron; D008875:Middle Aged; D008969:Molecular Sequence Data; D010802:Phylogeny; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D012321:DNA-Directed RNA Polymerases