Detection of Norwalk virus antibodies and antigen with a biotin-avidin immunoassay

Biotin-avidin immunoassays (BAIs) were developed to detect Norwalk virus antigen and to measure Norwalk virus antibody. The BAI detected Norwalk virus infections by a fourfold titer rise in antibody in sera or by antigen in stool, with a sensitivity similar to or greater than that of the radioimmunoassay (RIA), and the BAI appeared to be more sensitive than the RIA for detecting antibody in single serum specimens. The BAI antigen test detected Norwalk antigen in all stools that were positive by the RIA, and in two stools negative by the RIA. Of 106 serum pairs tested by both the RIA and BAI, 94 demonstrated seroconversion, or lack of seroconversion, in both assays; 12 gave discordant results. Titers by the BAI generally exceeded those by the RIA by two- or fourfold. The BAI had advantages over the RIA in that it had added sensitivity in the detection of Norwalk antibody, was less time consuming, did not require frequent labeling of antibody, and did not have the problems associated with the use of isotopes.