Cell culture propagation of rotaviruses

Abstract

Rotaviruses are ubiquitous intestinal pathogens that infect the young of many species. Their adaptation to cell culture has met with variable success depending on the strain and serogroup of virus. Successful cultivation of rotaviruses has been achieved through sequential use of animal passage and kidney cell cultures. Roller tubes were beneficial for adaption of several strains as opposed to use of stationary cell cultures. Activation of rotaviruses with proteolytic enzymes has increased their adaptability to culture and is standard practice with many strains. Alternative methods for cultivation of rotavirus, particularly human strains, have involved use of reassortants between animal and human rotaviruses. Continuous cell lines for rotavirus passage include rhesus monkey kidney (MA-104 & LLC-MK2), African green monkey kidney (BSC-1 & CV-1), and Madin-Darby bovine kidney. Fluorescent foci, cytopathic effects, and plaque assays are used to titrate infectious virus. Enzyme-linked immunosorbent assay, hemagglutination, dsRNA electropherotyping, and electron microscopy are used to detect rotavirus growth. Optimal conditions for the growth of two group A rotaviruses (OSU & NCDV) in MA-104 cells are described in this report.