Isolation and serial propagation of human group C rotaviruses in a cell line (CaCo-2)

✍ Scribed by Shinozaki, Kuniko; Yamanaka, Takaya; Tokieda, Masayoshi; Shirasawa, Hiroshi; Simizu, Bunsiti

• Publisher: John Wiley and Sons
• Year: 1996
• Tongue: English
• Weight: 603 KB
• Volume: 48
• Category: Article
• ISSN: 0146-6615
• DOI: 10.1002/(sici)1096-9071(199601)48:1<48::aid-jmv8>3.0.co;2-l

✦ Synopsis

Rotaviruses were detected via electron microscopy in fecal specimens collected from school children during an outbreak of diarrhea and from a sporadic case in 1993 in Japan. All of the viruses were found to belong to human group C rotavirus by reverse passive hemagglutination assay (RPHA). These viruses replicated well in a human colon carcinoma (CaCo-2) cell line cultured in the presence of trypsin (4 p,g/mI). This report demonstrates that human group C rotaviruses can be propagated efficiently i n a cell line cultured in the presence of trypsin. The infected cells did not show any apparent cytopathic changes. However, virus was detected in the cell cytoplasm by i m m u nof I uorescence (IF) staining and in the culture supernatant by RPHA. On the basis of immune electron microscopy (IEM), virus particles collected from infected CaCo-2 cell cultures were confirmed to aggregate specifically with anti-human group C rotavirus antibody. The electrophoretic patterns of RNA segments extracted from viral particles found in the fecal specimens or infected cells were identical to those of human group C rotavirus. These results indicated that human group C rotaviruses were the causal agent of the diarrhea outbreak.