Comparison of a whole-virus enzyme immunoassay (EIA) with a peptide-based EIA for detecting rubella virus immunoglobulin G antibodies following rubella vaccination

Abstract

A total of 250 human serum samples were tested for rubella virus immunoglobulin G antibodies by two enzyme immunoassays (EIAs), one using whole rubella virus antigen and the other based on the use of synthetic peptide antigen. The samples were taken from 125 volunteers before and after their immunization with the RA 27/3 rubella vaccine. This study indicates that a synthetic peptide-based EIA can favorably replace current viral lysate-based EIAs to detect rubella virus antibodies following immunization. Because the synthetic peptide used in this newly developed EIA represents a putative neutralization epitope of the rubella virus, it could also be instrumental in determining rubella immune status and in assessing vaccine program efficiency.

Rubella reimmunization: comparative analysis of the immunoglobulin G response to rubella virus vaccine in previously seronegative and seropositive individuals

“…a P values for comparisons between subjects whose sera exhibited a shift in immunoblot pattern from those subjects whose postvaccination sera exhibited no change in RV protein specificities as determined by IgG immunoblot assays. 239 (29,46), which is one of four known NT domains on this protein (8,40,43). GMTs and median antibody levels measured by the three whole-RV EIAs were determined to be well above their established negative cutoff points, suggesting that the majority of the individuals studied had protective levels of RV-specific IgG both before and after receiving the MMR vaccine.…”

“…This EIA uses as a target antigen a cyclized synthetic peptide, BCH-178c, representing E1 protein residues 213 to 239. Evidence that this sequence contains an NT domain includes the ability of synthetic peptides representing this E1 region (i) to bind to murine monoclonal antibodies with RV-neutralizing activity (8,29,43), (ii) to induce NT antibodies in mice and rabbits (13), and (iii) to bind to human sera containing RV NT antibodies (29,46). In the present study, it was observed that many individuals who would have been considered seropositive by the whole-RV EIAs were actually seronegative (titers, Ͻ10 IU/ml) by the E1 peptide EIA.…”

“…Synthetic peptide EIA. Specific IgG directed to an RV E1 protein NT domain were measured by a commercial EIA (DETECT-RUBELLA G; BioChem Immunosystems, Inc., Montreal, Quebec, Canada), which uses a cyclized synthetic peptide, BHC-178c (29,46) representing E1 residues 213 to 239. This region is conserved among all known strains of RV, including the HPV77/DE5 and RA27/3 vaccine strains.…”

“…Synthetic peptides have been used to map one NT domain located between E1 residues 213 and 239 recognized by both murine and human antibodies (29). Those investigations have indicated that after RV infection or vaccination, antibodies directed to E1 (residues 213 to 239) increase in parallel and correlate with the RV-specific antibodies measured by other EIAs and by HAI and NT tests (29,46). The E1 NT domain peptide-specific antibodies are lacking in individuals who remain seronegative, despite multiple RV vaccinations, who have been exposed to RV by congenital infection (29), or who show adverse reactivity to RV such as arthritis associated with persistent RV infection (27,28).…”

Presence of a neutralizing domain in isolates of rubella virus in Cordoba, Argentina

“…These data suggest that the RV epitopes contained by SP15 may be important for protective host humoral immune responses. Moreover, other authors (10,22) reported that a similar sequence (E1 213-239 ), represented by peptide BCH-178, defines a neutralization epitope on RV for human antibodies and is useful in determining RV immunity. The critical sequences of synthetic peptides required for antibody induction often lie in regions of a virus that are quite variable among different isolates and strains.…”