Radioimmunoassay for detection of antibodies to Epstein-Barr virus in human infectious mononucleosis serum specimens

Abstract

A rapid microradioimmunoassay (RIA) technique was adapted for quantitatively measuring antibody titers to antigens occurring in Epstein-Barr virus (EBV)-infected lymphoid cells. In these experiments two EBV-infected cell lines, HR1K and EB-3, were used as antigen-positive cells and Molt-4 was used as the negative control cells. The antibody titers of sera from suspected infectious mononucleosis patients were compared by RIA and indirect fluorescent antibody (IFA) methods. As determined by each of the methods, 14 of 19 sera had positive antibody titers and the remainder of the sera had negative antibody titers. Thus, the two methods agreed completely in differentiating sera with antibodies to EBV antigens. To further evaluate the antibody specificity of the RIA, the antibody titers of paired sera, pre- or early infection and postinfection, from five confirmed infectious mononucleosis patients were determined by RIA and IFA. Seroconversion was demonstrated by both RIA and IFA for each of the patients. Thus, the sensitivity and specificity of the two procedures are about the same.

Detection of virus-specific immunoglobulins using a doubly labeled fluorescein-125I antibody

“…The titers of both virus-specific immunoglobulin and virus-specific IgG obtained by RIA match closely those of the IFA method. This is consistent with results obtained by other workers (6).…”

“…Efforts are being made to develop a reliable radioimmunoassay (RIA) for the detection of virus-specific immunoglobulins, acceptable for use in diagnostic serology (1,2,(5)(6)(7)(8). The establishment of a satisfactory RIA depends on the use of antibody with both a high avidity and selectivity for the material to be assayed (4).…”

Sensitivity of a radioimmunoassay method for detection of certain viral antibodies in sera and cerebrospinal fluids

“…These studies demonstrate the applicability of an indirect solid-phase RIA system, utilizing fixed virus-infected cells, for titration of serum and CSF antibodies to a variety of human viruses, and they show the RIA system to be markedly more sensitive, in terms of antibody titers detected, than conventional serological tests used for assay of viral antibodies. They also indicate the appropriate dilutions at which Antibody titers demonstrated in our RIA system were markedly higher than those reported by certain other investigators who used lyates of virus-infected cells adsorbed onto plastic surfaces as a source of viral antigen, and found antibody titers demonstrated by RIA to be only slightly higher than those obtained in conventional viral antibody assays (10,12,27). These differences in sensitivity may be due in part to differences in labeling methods (11) and in part to the fact that fixed virus-infected cells, as first utilized by Hayashi et al (7) for RIA of HSV antibodies, provide a more suitable source of antigen.…”

“…Since other viral antigens do not possess these characteristics, development of sensitive RIA techniques has been more difficult. Some of the RIA methods described for assay of viral antibodies have had little or no greater sensitivity, from the standpoint of antibody titers demonstrated, than conventional viral antibody assay methods such as complement fixation (CF), hemagglutination inhibition (HI), neutralization, or fluorescent-antibody (FA) staining (10,12,27), and thus there has been no particular advantage to the diagnostic laboratory in adopting these methods, which require unstable, and potentially hazardous, radioisotopes and special equipment for counting isotope emissions.…”

Local antibody response in chickens: analysis of antibody synthesis to Newcastle disease virus by solid-phase radioimmunoassay and immunofluorescence with class-specific antibody for chicken immunoglobulins

“…Radioimmunoassay. The solid-phase radioimmunoassay described previously (7,(14)(15)(16)32) was modified for quantitation of class-specific anti-NDV immunoglobulins in chickens (technique to be reported elsewhere). Briefly, stocks of NDV antigen were prepared in cell culture.…”