Use of antisera against bovine (NCDV) and simian (SA11) rotaviruses in ELISA to detect different types of human rotavirus

Abstract

Two ELISA systems for the detection of human rotaviruses were developed. In the first system antibodies to Nebraska calf diarrhea virus (NCDV) were used for coating the solid matrix and for the preparation of the enzyme conjugate. In the second system antibodies to human rotavirus and antibodies to simian rotavirus (SA11) were used for coating the solid matrix and for the preparation of the enzyme conjugate respectively. The second ELISA system proved to have a broader spectrum for the detection of human rotaviruses. By using the two ELISA systems, the different types of human rotavirus could be distinguished. The ELISA tests developed were 8 to 64 times as sensitive as electron microscopy (EM) and (or) counterimmunoelectrophoresis (CIEP). The antigen detected by ELISA was shown to be different from that detected by the hemagglutination test.

Detection of respiratory syncytial, parainfluenza type 2, and adenovirus antigens by radioimmunoassay and enzyme immunoassay on nasopharyngeal specimens from children with acute respiratory disease

“…The mucus fractions were obtained from 8, the cell fractions were obtained from 6, and the original nasopharyngeal secretion was available from 3 of the 11 IF-positive specimens. All specimen fractions for the 11 IF-positive specimens were positive, and all IF-negative specimens were negative in adenovirus RIA, and the counts-per-minute values were comparable to those obtained in RSV and parainfluenza type 2 RIAs (Table 4).…”

“…Radioimmunoassay (RIA) and enzyme immunoassay (EIA) methods recently have been applied in the direct detection of viral antigens in clinical specimens (2,6,9,10,12,13). These methods offer high sensitivity and specificity, do not require any laborious pretreatment of the specimens, and allow the testing of a large number of specimens per day.…”

Human rotavirus antigen detection by enzyme-immunoassay with antisera against Nebraska calf diarrhoea virus.

“…Bishai et al,17 on the other hand, found several electron microscopically-positive specimens negative by EIA using NCDV antisera as immunoreagents but positive by EIA using human rotavirus and simian rotavirus (SAI 1) antisera as immunoreagents. The discrepancy between the results of Bishai et al 17 and the results of this study might be explained by the fact that they used complete virus particles for the production of immune sera whereas incomplete virus particles were used in the present study.…”

Antigenic characterization of human and animal rotaviruses by immune adherence hemagglutination assay (IAHA): evidence for distinctness of IAHA and neutralization antigens

“…Finally, with selective hyperimmune animal sera prepared against two strains of complete human rotavirus virions, i.e., virus that possessed a double capsid, two antigenically distinct human rotavirus groups, designated serotype 1 and serotype 2, were detected by CF, immune electron microscopy, and ELISA (3,(33)(34)(35). The existence of at least two human rotavirus serotypes was also demonstrated by fluorescent focus neutralization with human convalescent sera or animal hyperimmune rotavirus antisera and was also suggested by a binding ELISA with hyperimmune rotavirus antisera (1,2,6,25).…”

“…This property made it possible to use cultivatable animal rotaviruses as substitute antigens for detection of serological evidence of infection with the fastidious, non-cultivatable human rotaviruses (12). Later, more specific antigens were detected by neutralization of abortive foci of infection in tissue culture (fluorescent focus neutralization), hemagglutination inhibition, and enzyme-linked immunosorbent assay (ELISA) blocking and binding techniques which demonstrated that human rotaviruses differed antigenically from ani-41E mal rotaviruses and that animal rotaviruses differed from each other as well (2,22,26,32). Finally, with selective hyperimmune animal sera prepared against two strains of complete human rotavirus virions, i.e., virus that possessed a double capsid, two antigenically distinct human rotavirus groups, designated serotype 1 and serotype 2, were detected by CF, immune electron microscopy, and ELISA (3,(33)(34)(35).…”