Simple and specific enzyme immunoassay using monoclonal antibodies for serotyping human rotaviruses

Publisher Website
Google Scholar
Abstract
An enzyme immunoassay for serotyping human rotaviruses in stools and in cell culture was developed. Hyperimmune rabbit antisera to rotaviruses were used as capture antibodies, and rotavirus-neutralizing mouse monoclonal antibodies specific for serotypes 1, 2, 3, and 4 were used as detection reagents. Partial purification of monoclonal antibodies and inclusion of skim milk powder of antibody diluents contributed to assay specificity. The senstivity of this assay was greater than that of a direct enzyme immunoassay in which rotaviruses of the appropriate serotype were absorbed directly to the solid phase. When fecal extracts were concentrated threefold, this serotyping enzyme immunoassay was of equal specificity and approached the sensitivity of electron microscopy for rotavirus detection. This assay is simple and rapid and is suitable for serotyping the large numbers of isolates obtained from epidemiological studies and vaccine trials.