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Immunosorbent assay based on recombinant hemagglutinin protein produced in a high-efficiency mammalian expression system for surveillance of measles immunity.
J Clin Microbiol. 1998 Mar; 36(3):721-6.JC

Abstract

Recombinant hemagglutinin (H) protein of the measles virus (MV) was produced in mammalian cells with a high-yield expression system based on the Semliki Forest virus replicon. Crude membrane preparations of H protein-transfected BHK-21 cells were used to coat microtiter plates to measure specific immunoglobulin G antibodies in 228 serologically defined serum samples mainly from measles late-convalescent adults. The titers by the enzyme-linked immunosorbent assay for the H protein (H-ELISA) closely correlated with neutralization test (NT) titers (R2 = 0.66), hemagglutination inhibition test (HI) titers (R2 = 0.64), with the titers from a certified commercial ELISA based on whole MV-infected cells (MV-ELISA; R2 = 0.45). The correlations described above were better than those of the commercial MV-ELISA titers with the NT (R2 = 0.52) or HI (R2 = 0.48) titers. By using the 2nd International Standard for anti-measles serum, the detection level of the assay corresponds to 215 mIU/ml for undiluted serum, which corresponds to the estimated threshold for protective immunity. The specificity, accuracy, and positive predictive value were, in general, better for the H-ELISA than for a commercial MV-ELISA, independent of whether HI, NT, or HI and NT were used as "gold standards." In contrast, the H-ELISA proved to be slightly less sensitive than the MV-ELISA (sensitivities, 98.6 versus 99.5%, respectively; P was not significant). The assays did not differ significantly in the number of serum samples with positive HI and NT results (n = 212) which measured false negative (H-ELISA, 2 of 212 [0.94%]; MV-ELISA, 1 of 212 [0.47%]), but the H-ELISA detected significantly more measles-susceptible individuals than the MV-ELISA (10 of 11 versus 3 of 11, respectively; P < 0.05) among the individuals whose sera had negative HI and NT results. Our data demonstrate that the H-protein preparation that we describe could be a cost-effective alternative to current whole-virus-based ELISAs for surveillance for immunity to measles and that such an assay could be more efficient in detecting susceptibility to measles. Furthermore, unlike whole MV-based antigens, H-protein would also be suitable for use in the development of a simple field test for the diagnosis of measles.

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Authors+Show Affiliations

Bouche F
Laboratoire National de Santé, Luxembourg, Luxembourg.
Ammerlaan W
No affiliation info available
Berthet F
No affiliation info available
Houard S
No affiliation info available
Schneider F
No affiliation info available
Muller CP
No affiliation info available

MeSH

AdolescentAdultAnimalsAntibodies, ViralCell LineEnzyme-Linked Immunosorbent AssayFalse Negative ReactionsFalse Positive ReactionsFemaleHemagglutinins, ViralHumansImmunoglobulin GInfantMaleMeaslesMeasles virusPredictive Value of TestsRecombinant ProteinsSensitivity and SpecificityTransfection

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

9508302

Citation

Bouche, F, et al. "Immunosorbent Assay Based On Recombinant Hemagglutinin Protein Produced in a High-efficiency Mammalian Expression System for Surveillance of Measles Immunity." Journal of Clinical Microbiology, vol. 36, no. 3, 1998, pp. 721-6.
Bouche F, Ammerlaan W, Berthet F, et al. Immunosorbent assay based on recombinant hemagglutinin protein produced in a high-efficiency mammalian expression system for surveillance of measles immunity. J Clin Microbiol. 1998;36(3):721-6.
Bouche, F., Ammerlaan, W., Berthet, F., Houard, S., Schneider, F., & Muller, C. P. (1998). Immunosorbent assay based on recombinant hemagglutinin protein produced in a high-efficiency mammalian expression system for surveillance of measles immunity. Journal of Clinical Microbiology, 36(3), 721-6.
Bouche F, et al. Immunosorbent Assay Based On Recombinant Hemagglutinin Protein Produced in a High-efficiency Mammalian Expression System for Surveillance of Measles Immunity. J Clin Microbiol. 1998;36(3):721-6. PubMed PMID: 9508302.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Immunosorbent assay based on recombinant hemagglutinin protein produced in a high-efficiency mammalian expression system for surveillance of measles immunity. AU - Bouche,F, AU - Ammerlaan,W, AU - Berthet,F, AU - Houard,S, AU - Schneider,F, AU - Muller,C P, PY - 1998/3/21/pubmed PY - 1998/3/21/medline PY - 1998/3/21/entrez SP - 721 EP - 6 JF - Journal of clinical microbiology JO - J Clin Microbiol VL - 36 IS - 3 N2 - Recombinant hemagglutinin (H) protein of the measles virus (MV) was produced in mammalian cells with a high-yield expression system based on the Semliki Forest virus replicon. Crude membrane preparations of H protein-transfected BHK-21 cells were used to coat microtiter plates to measure specific immunoglobulin G antibodies in 228 serologically defined serum samples mainly from measles late-convalescent adults. The titers by the enzyme-linked immunosorbent assay for the H protein (H-ELISA) closely correlated with neutralization test (NT) titers (R2 = 0.66), hemagglutination inhibition test (HI) titers (R2 = 0.64), with the titers from a certified commercial ELISA based on whole MV-infected cells (MV-ELISA; R2 = 0.45). The correlations described above were better than those of the commercial MV-ELISA titers with the NT (R2 = 0.52) or HI (R2 = 0.48) titers. By using the 2nd International Standard for anti-measles serum, the detection level of the assay corresponds to 215 mIU/ml for undiluted serum, which corresponds to the estimated threshold for protective immunity. The specificity, accuracy, and positive predictive value were, in general, better for the H-ELISA than for a commercial MV-ELISA, independent of whether HI, NT, or HI and NT were used as "gold standards." In contrast, the H-ELISA proved to be slightly less sensitive than the MV-ELISA (sensitivities, 98.6 versus 99.5%, respectively; P was not significant). The assays did not differ significantly in the number of serum samples with positive HI and NT results (n = 212) which measured false negative (H-ELISA, 2 of 212 [0.94%]; MV-ELISA, 1 of 212 [0.47%]), but the H-ELISA detected significantly more measles-susceptible individuals than the MV-ELISA (10 of 11 versus 3 of 11, respectively; P < 0.05) among the individuals whose sera had negative HI and NT results. Our data demonstrate that the H-protein preparation that we describe could be a cost-effective alternative to current whole-virus-based ELISAs for surveillance for immunity to measles and that such an assay could be more efficient in detecting susceptibility to measles. Furthermore, unlike whole MV-based antigens, H-protein would also be suitable for use in the development of a simple field test for the diagnosis of measles. SN - 0095-1137 UR - https://www.unboundmedicine.com/medline/citation/9508302/Immunosorbent_assay_based_on_recombinant_hemagglutinin_protein_produced_in_a_high_efficiency_mammalian_expression_system_for_surveillance_of_measles_immunity_ L2 - http://jcm.asm.org/cgi/pmidlookup?view=long&amp;pmid=9508302 DB - PRIME DP - Unbound Medicine ER -