R G Wyatt

Enzyme-linked immunosorbent assay (ELISA) for detection of human reovirus-like agent of infantile gastroenteritis.

An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of the human reovirus-like agent of infantile gastroenteritis in human stools. The results of the assay can be read either with a simple colorimeter or the naked eye. Investigations with 143 samples from children with gastroenteritis and 75 samples from children with other illnesses showed that the ELISA was as sensitive as electron microscopy or radioimmunoassay for detection of this agent. In addition, the ELISA was simple to perform and, when read visually, did not require sophisticated technical equipment. These advantages make it suitable for field work.

Epidemiology of human rotavirus Types 1 and 2 as studied by enzyme-linked immunosorbent assay.

To determine the relative importance of two known serotypes of human rotavirus, we developed an enzyme-linked immunosorbent assay to differentiate serotype-specific rotavirus antigen and antibody. Using this technic, we studied the epidemiology of the two serotypes in acute gastroenteritis. Seventy-seven per cent of 414 rotavirus isolates were Type 2, and the remainder were Type 1. The serotype distribution was similar in specimens from children in Washington, D.C., and other parts of the world. Sero-epidemiologic studies revealed that most children living in the Washington, D.C., area acquired antibody to both types by the age of two years. An analysis of children who were reinfected indicated that sequential infections usually involved different serotypes and that illness caused by one serotype did not provide resistance to illness caused by the other serotype. These results suggest that, to be completely effective, a vaccine must provide resistance to both serotypes.

NEW COMPLEMENT-FIXATION TEST FOR THE HUMAN REOVIRUS-LIKE AGENT OF INFANTILE GASTROENTERITIS: Nebraska Calf Diarrhœa Virus Used as Antigen

A complement-fixation (C.F.) test for the human reovirus-like agent of infantile gastroenteritis has been developed using the serologically related Nebraska calf diarrhoea virus (N.C.D.V.) as antigen. Most infants and children who shed the agent in stools and/or who demonstrated serological (C.F.) evidence of infection with a reovirus-like-particle-positive human stool-filtrate C.F. antigen also demonstrated serological evidence of infection when a concentrated N.C.D.V. preparation was employed AS C.F. antigen. The N.C.D.V., which was previously shown to be related to the human reovirus-like agent, was found to be related antigenically to the epizootic diarrhoea of infant mice (E.D.I.M.) virus also. Studies on the prevalence of C.F. antibody in sera from infants and young children revealed a pattern of rapid acquisition of antibody to both the human reovirus-like agent and the N.C.D.V. as over 80 percent of these individuals possessed antibody to each agent by 36 months of age. A strong positive association was found in the results obtained with the two antigens. The ready availability of cell-culture grown N.C.D.V., and its ability to serve as a substitute C.F. antigen for the human reovirus-like agent, should enable the serodiagnosis of many cases of disease due to the human agent and facilitate seroepidemiological studies of such infections. In addition, the observation that a large proportion of individuals infected with the human reovirus-like agent develop serological evidence of infection not only to the human agent but to the calf agent as well may have important implications in the immunoprophylaxis of disease caused by the human reovirus-like agent.

A DOT HYBRIDISATION ASSAY FOR DETECTION OF ROTAVIRUS

A dot hybridisation technique for the detection of rotavirus in stools and other biological materials is described. The assay is based on the in-situ hybridisation of labelled single-stranded RNA probes, obtained by in-vitro transcription of rotavirus particles, to heat-denatured rotavirus RNA immobilised on nitrocellulose membranes. The method is highly specific and allows for the detection of as little as 8 pg of viral RNA. Its use for the detection of rotavirus in stool suspensions and rectal swabs obtained from children with diarrhoea may facilitate epidemiological studies of rotavirus gastroenteritis.