POLYACRYLAMIDE GEL ELECTROPHORESIS (PAGE) OF ROTAVIRUS RNA AFTER SERIAL PASSAGE AND IN CHRONIC INFECTION

Abstract

Rotaviruses are important causes of serious gastroenteritis in infants and children. New strains of rotavirus emerge during the course of an epidemic, but little is known about the mechanisms by which this occurrs. It is not known, for example, if significant alterations occur in the rotavirus genome during direct transmission from one individual to another. We investigated this question through the analysis of the distinctive electropherotypes produced by high resolution PAGE of rotavirus dsRNA from an animal model and from children with severe combined immunodeficiency disease (SCID) who had prolonged excretion of rotavirus.To test the stability of the rotavirus genome during multiple infections, a strain of murine rotavirus was serially transmitted for more than 10 passages over a period of several months. No changes in the rotavirus electropherotype occurred during this period of infection. In the case of human infection, rotavirus was isolated from multiple stool specimens collected from two SCID patients over the course of several weeks and analyzed for RNA patterns. Each specimen contained more than one strain of rotavirus as determined by examination of the PAGE electropherotype. Marked changes were noted in rotavirus electropherotypes from different specimens from each patient. Recombinant events or recurrent infection with multiple rotavirus strains might account for the appearance of new strains of human rotavirus and might contribute to the chronic diarrhea in such patients.