3'-terminal sequence of a small round structured virus (SRSV) in Japan.
We determined the nucleotide sequence of about 1,000 bases from the 3'-terminus of a small round structured virus (SRSV), which caused a gastroenteritis outbreak in Chiba Prefecture, Japan, in 1987. The sequence was compared with the corresponding sequence region of Norwalk virus; it consisted of a part of the open reading frame 2 (ORF2), whole ORF3, and 3'-noncoding region (NCR). The 624-base-long ORF3 had sequence homology of 68% with the corresponding region of Norwalk virus. (The amino acid sequence homology was 74%.) The 94-base-long NCR had 65% homology with Norwalk virus. We then selected two consensus-sequence portions in the above sequence between Chiba and Norwalk viruses for primers in the reverse transcriptase-polymerase chain reaction (RT-PCR). Using this primer set, we detected 669-bp bands in agarose gel electrophoresis of RT-PCR products from feces containing Chiba or Norwalk viruses. Furthermore, in Southern hybridization with Chiba probes which were labeled with digoxigenin-dUTP in PCR, the bands of the two viruses were clearly stained under a low stringency condition. Since both Chiba and Norwalk viruses were detected by the above primer set although they are geographically and chronologically different viruses, our primer-pair may be useful for detection of a broad range of SRSVs which cause gastroenteritis in different areas.
Utagawa
E T
ET
Department of Epidemiology, National Institute of Health, Tokyo, Japan.
Takeda
N
N
Inouye
S
S
Kasuga
K
K
Yamazaki
S
S
D001483:Base Sequence; D002139:Caliciviridae / Q000235:genetics* / Q000302:isolation & purification; D017931:DNA Primers; D005759:Gastroenteritis / Q000382:microbiology*; D006801:Humans; D007564:Japan; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid
D017931:DNA Primers