Evaluation of nine sets of PCR primers in the RNA dependent RNA polymerase region for detection and differentiation of members of the family Caliciviridae, Norwalk virus and Sapporo virus.
Norwalk virus and Sapporo virus were approved as type species of the genus "Norwalk-like viruses" and the genus "Sapporo-like viruses," respectively, in the family Caliciviridae. A total of 116 stool specimens containing Norwalk virus (NV) or Sapporo virus (SV) were tested by RT-PCR and Southern hybridization to evaluate nine sets of PCR primers and seven internal oligonucleotide probes in the RNA dependent RNA polymerase region of NV and SV for detection and differentiation of viruses in the NV and SV. Fifty-five stool samples were collected from 11 outbreaks of NV and/or SV gastroenteritis in an infant home, where residents were infants under 2 years of age, in Sapporo, Japan. Sixty specimens were obtained in Sapporo from sporadic cases in children, mainly under 6 years of age, of acute gastroenteritis due to small round structured viruses detected by EM. There is no single primer pair to detect all NV and SV, and at least three primer pairs, G1 set, G2 set and Sapp35/Sapp36, are required to detect viruses in the NV and SV clades. Many NV and SV strains were successfully classified into one of the NV/genogroup I, NV/genogroup II and SV by single-round RT-PCR and Southern hybridization. The new detection method for SV reported in this study combined with those for NV previously reported may elucidate the importance of Norwalk virus and Sapporo virus as a cause of viral gastroenteritis in all age groups in the world.
Honma
S
S
Department of Pediatrics, Sapporo Medical University School of Medicine, Hokkaido, Japan.
Nakata
S
S
Kinoshita-Numata
K
K
Kogawa
K
K
Chiba
S
S
D015139:Blotting, Southern; D002139:Caliciviridae / Q000145:classification* / Q000235:genetics; D017250:Caliciviridae Infections / Q000821:virology; D002675:Child, Preschool; D017931:DNA Primers / Q000138:chemical synthesis; D005243:Feces / Q000821:virology; D005759:Gastroenteritis / Q000821:virology; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D009663:Norwalk virus / Q000145:classification* / Q000235:genetics; D015345:Oligonucleotide Probes; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D012324:RNA-Dependent RNA Polymerase / Q000032:analysis / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D017931:DNA Primers; D015345:Oligonucleotide Probes; D012367:RNA, Viral; D012324:RNA-Dependent RNA Polymerase