[The detection of infectious rotaviruses by cell culture technique: use and evaluation of the immunoperoxidase assay]

Abstract

In the present study, virus isolation was attempted in one hundred and twenty-one fecal samples of children suffering from acute gastroenteritis. Virus isolation was performed either conventionally by examination of cytopathogenic effect (CPE) or by immunoperoxidase staining (IPS) of rotavirus group specific antigen (inner capsid) in trypsin free MA104 cells within 18 h. Applying the conventional technique, rotavirus was isolated in only 4 (3.3%) fecal specimens. In contrast, IPS detected infectious virus in 49 (40.5%) stool samples. The specificity of IPS was confirmed by the results obtained with an antigen detection ELISA ("Rotazyme", Abbott, Wiesbaden) and gel electrophoresis of rotaviral RNA (electrophoretyping). ELISA and RNA gel electrophoresis detected rotavirus in 66 (54.4%) and 56 (46.3%) stool samples respectively. IPS enables rapid diagnosis of rotavirus infections in cell cultures applied to detect infectious viral particles in order to be used in the investigation of nosocomial outbreaks, material- and surface contamination and evaluation of disinfectants.