Hepatitis A virus was purified from fecal specimens obtained from 3 patients with naturally acquired hepatitis A, by a process of differential centrifugation, chloroform extraction, column chromatography, and isopycnic ultracentrifugation. Analysis of purified virus by discontinuous SDS-PAGE revealed three major polypeptides with molecular weights of 34,000, 25,500, and 23,000 daltons. These polypeptides appear to be specific for hepatitis A virus and have similar molecular weights to three of the four major polypeptides reported for members of the genus Enterovirus within the family Picornaviridae.

No abstract

Three monoclonal antibodies (K2-4F2, K3-2F2, and K3-4C8) of the immunoglobulin G2a class were raised against hepatitis A virus. The specificity of these antibodies was confirmed by immune electron microscopy, solid-phase radioimmunoassay, and in vitro neutralization in cell culture. Binding studies suggested that they all recognize closely related antigenic determinants. These monoclonal antibodies should prove to be of great value as diagnostic and research reagents.

Hepatitis A virus was purified from fecal specimens obtained from 2 patients with naturally acquired hepatitis A. The purification procedure involved differential centrifugation, organic solvent extraction, agarose gel filtration, ion-exchange chromatography, and isopycnic ultracentrifugation in cesium chloride. Using immune electron microscopy and discontinuous SDS-PAGE, this procedure was found to be effective in removing extraneous material from hepatitis A virus. There was significant recovery of virus as judged by immune electron microscopy and solid-phase radioimmunoassay. Using this protocol, it has been possible to obtain virus preparations of sufficient purity and high enough titer to enable biochemical studies to proceed.

A prospective study was carried out on 200 patients admitted to Fairfield Hospital, Melbourne, Australia, with acute hepatitis A to determine the frequency with which virus could be detected in their feces. Evidence of infection with hepatitis A virus (HAV) was obtained by detecting IgM specific for HAV in a single serum sample or by demonstrating a rising titer of antibody in paired sera by solid-phase radioimmunoassay. HAV was detected in the feces of 59 of the 200 patients by solid-phase radioimmunoassay and immune electron microscopy. When patients were admitted within one week of the onset of dark urine, 45% were found to be shedding HAV, whereas only 11% of specimens obtained from patients admitted during the second week contained virus. HAV was not detected in fecal specimens collected more than 14 days after the onset of dark urine. These findings suggest that patients admitted to hospital with hepatitis A may still be infectious and that appropriate precautions against fecal contamination should be maintained.