WRRCTR No.161 Replication of Human Rotavirus in Tissue Culture: Recovery and Detection in Fecal, Sewage, and Natural Water Samples

Human rotavirus is the major cause of gastroenteritis among young children. To replicate this virus, sensitive methods using standard tissue culture systems are required. The project goal was to develop laboratory capability to recover and detect this infectious rotavirus in fecal, sewage, and natural water samples. Using simian rotavirus (SA-II) as a model system and an enzyme-linked immunosorption (ELISA) test capable of detecting high concentrations of rotavirus, the protamine sulfate method was determined as superior to the aluminum chloride precipitation and polymer two-phase methods for recovering rotavirus from sewage. The ELISA method was very effective in detecting rotavirus in stool samples of children. Stools from children not displaying clinical symptoms of rotavirus infection were negative for rotavirus, whereas 43 to 58% of stools from children displaying clinical symptoms of rotavirus infection was positive for rotavirus. The results suggested an association of increased rotavirus infections during the winter months in the state of Hawaii. Stools positive for rotavirus by the ELISA test were used as inoculum to develop methods to replicate human rotavirus in the cell culture system. After many unsuccessful attempts, human rotavirus was cultured after two passages in primary cynomolgus monkey kidney cells. Human rotavirus which replicated in the primary monkey kidney cells was shown to be capable of replicating in continuous monkey kidney cell lines such as the MA-104.U.S. Department of the Interior Grant/Contract No. CT371300; 37130