Evaluation of Hemagglutinin Protein-Specific Immunoglobulin M for Diagnosis of Measles by an Enzyme-Linked Immunosorbent Assay Based on Recombinant Protein Produced in a High-Efficiency Mammalian Expression System

Abstract

Recombinant hemagglutinin (H) of the measles virus (MV) expressed in a mammalian high-expression system based on the Semliki Forest virus replicon was used in an enzyme-linked immunosorbent assay (ELISA) for the detection of specific immunoglobulin M (IgM) and IgG in patients with acute-phase measles. One hundred twelve serum specimens from 70 patients with measles were analyzed. Case definition was based on a commercial IgM ELISA that utilizes MV-infected cells (MV-ELISA) (Enzygnost; Behring Diagnostics); the clinical criteria of the Centers for Disease Control and Prevention (Atlanta, Ga.); and/or the increase in hemagglutinin test titers, neutralization test titers, and levels of MV-specific IgG whenever paired sera were available. The initial time courses of the IgM signal after the onset of rash are similar in the H- and MV-ELISAs. On days 0 to 19, both ELISAs detected IgM in 67 of 68 (98.5%) sera. Average maximal levels of IgM seem to persist, however, about 10 days longer in the MV-ELISA (up to day 25) than in the H-ELISA (day 15). From days 20 to 29 and 30 to 59, the H-ELISA detected only 64.3 (9 of 14) and 19.2% (5 of 26), respectively, of sera that were IgM positive by MV-ELISA. At least up to day 30, the performance of the H-ELISA seemed to be similar to that reported for commercial ELISAs based on whole MV. Our results demonstrate that MV H-specific IgM can be used to diagnose most measles cases from a single serum specimen collected within 19 days after the onset of rash and that the recombinant protein used in this study is suitable for this purpose.

Resistance of recent measles virus wild-type isolates to antibody-mediated neutralization by vaccinees with antibody

“…The isolates from Luxembourg (MV201, MV203, MV204, MV216, MV224, MV225, MV227) have been described by Hanses et al [2000]. MV201 and MV 216 were from an outbreak in which about 10% of cases were vaccinated previously [Mossong et al, 2000;Bouche et al, 1998]. Most viruses were isolated originally on B95a cells, but some viruses were passaged initially on Vero cells.…”

Measles virus protein‐specific IgM, IgA, and IgG subclass responses during the acute and convalescent phase of infection

“…Virus neutralising antibodies are directed towards the H and F transmembrane glycoproteins [Bouche et al, 2002]. The introduction of recombinant technology has enabled the detection of protein-specific responses at the isotype level [Beauverger et al, 1993;Bouche et al, 1998;De Swart et al, 1998;Hartter et al, 2000]. For the glycoprotein-specific responses, immunofluorescence assays using intact transfected target cells may be preferred since this results in detection of antibodies that recognise the extra-cellular domain of the protein in its natural conformation.…”