TY - JOUR
T1 - A simplified immunoassay based on measles virus recombinant hemagglutinin protein for testing the immune status of vaccinees
AU - Bouche, Fabienne
AU - Ammerlaan, Wim
AU - Fournier, Phillipe
AU - Schneider, François
AU - Muller, Claude P.
N1 - Funding Information:
We thank the children, their parents and teachers for the good collaboration. We are grateful to the Ministère de l'Education Nationale for permitting systematic blood collections in several schools, and we acknowledge the support of the Direction de la Santé of the Ministere de la Santé (Luxembourg). Parts of this work were done by F.B. in partial fulfillment of her doctoral thesis. This study was supported by a grant of the Centre de Recherche Public-Santé, Luxembourg (CRP93/08) and a bourse formation recherche to F.B.
PY - 1998/9
Y1 - 1998/9
N2 - 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. Copyright (C) 1998 Elsevier Science B.V.
AB - 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. Copyright (C) 1998 Elsevier Science B.V.
KW - ELISA
KW - Hemagglutination inhibition
KW - Hemagglutinin protein
KW - IgG
KW - Measles virus
KW - Neutralisation assay
KW - Vaccinees
UR - http://www.scopus.com/inward/record.url?scp=0031811906&partnerID=8YFLogxK
U2 - 10.1016/S0166-0934(98)00073-1
DO - 10.1016/S0166-0934(98)00073-1
M3 - Article
C2 - 9763131
AN - SCOPUS:0031811906
SN - 0166-0934
VL - 74
SP - 77
EP - 87
JO - Journal of Virological Methods
JF - Journal of Virological Methods
IS - 1
ER -