A simplified immunoassay based on measles virus recombinant hemagglutinin protein for testing the immune status of vaccinees.
Simplified tests based on recombinant antigens are considered to be important for monitoring immunity against measles virus (MV). The hemagglutinin protein (H) is the main target for neutralising and protective antibodies. We produced a recombinant MV-H protein, in a high-yield mammalian expression system based on the Semliki Forest virus replicon. The antigenicity of this recombinant protein was investigated with monoclonal antibodies and its suitability for measuring the immune status of vaccinees was tested in a large cohort by ELISA (H-ELISA). The results were evaluated against neutralisation (NT) and hemagglutination inhibition (HI) titers and MV-specific IgG measured in a commercial whole-virus based ELISA (MV-ELISA, Enzygnost). The H-ELISA correlated better with HI (r=0.78) and NT titers (r=0.80), than the MV-ELISA (HI, r=0.58; NT, r=0.59). In contrast to the MV-ELISA, the H-ELISA detected no false-positive sera (P < 0.02) and the number of false-negative sera was significantly lower in the H-ELISA than in the MV-ELISA (4/378 vs. 15/378; P < 0.025). The performance of the H-ELISA did not deteriorate significantly when, instead of background corrected net values, uncorrected raw O.D. values of the H-antigen were considered, or when early time points (30 min) were evaluated. These results demonstrate that the recombinant H-ELISA detects efficiently non-immune individuals among vaccinees, despite their relatively low MV-antibody levels. A simplified format with single value measurements did not result in loss of sensitivity or specificity and its performance compared favorably with commercial ELISAs based on whole virus.
Department of Immunology, Laboratoire National de Santé, Luxembourg, Luxembourg.
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F
Bouche
W
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Ammerlaan
P
P
Fournier
F
F
Schneider
C P
CP
Muller
D000293:Adolescent; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D002460:Cell Line; D002648:Child; D003124:Colorimetry; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006385:Hemagglutination Inhibition Tests; D006389:Hemagglutinins, Viral / Q000276:immunology*; D006801:Humans; D007118:Immunoassay; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D008297:Male; D008458:Measles Vaccine / Q000276:immunology*; D008459:Measles virus / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology
D000914:Antibodies, Viral; D006389:Hemagglutinins, Viral; D007074:Immunoglobulin G; D008458:Measles Vaccine; D011993:Recombinant Fusion Proteins; C032397:hemagglutinin protein G, measles virus