Development and Evaluation of a Capture ELISA for IgM Antibody to the Human Cytomegalovirus Major DNA Binding Protein

J Virol Methods. 1991 Dec;35(3):315-29. doi: 10.1016/0166-0934(91)90073-9.

Abstract

A new capture ELISA (ELAb) for determination of the IgM antibody response to the human cytomegalovirus major DNA binding protein (p52) was developed by using a p52-specific monoclonal antibody. As a reference test, a capture ELISA using in parallel viral- and cell-control labeled antigens (ELA) was employed. General specificity, which was determined on 180 unselected IgM-negative sera from an adult population was 100%; stringent specificity, which was evaluated on 108 potentially interfering sera from patients with Epstein-Barr virus infectious mononucleosis, autoimmune diseases, rheumatoid factor or treated with radioimmunotherapy, was 96.3%; finally, clinical specificity, determined on 79 IgM-negative sera drawn prior to onset of primary HCMV infection, was 100%. Thus, the overall specificity was 98.9% (363/367 IgM negative tested sera). Sensitivity assayed on 277 IgM-positive sera was 100%. The study of the kinetics of the IgM antibody response in sequential blood samples from 9 immunocompetent and 9 heart transplanted patients showed that, while in the immunocompetent p52-specific IgM titer fell sharply 2-3 months after onset and was virtually undetectable 12 months after onset, in the immunocompromised the IgM response persisted for longer than a year. Recurrent HCMV infections were associated with a high titer IgM response in 6 (30%), and with a low IgM response in another 6 (30%) heart transplanted patients within a group of 20 patients sequentially examined. Finally, IgM antibodies were detected in all 4 infants with congenital infection and in 5 of 6 infants with neonatal infection. The results show that the HCMV p52-specific IgM antibody response parallels that obtained by ELA, thus representing a major component of it. ELAb is highly sensitive, specific and reproducible. It represents a major advance among capture ELISA techniques, allowing detection of IgM antibody reactive to a specific viral protein.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal
  • Antibodies, Viral / immunology*
  • Autoimmune Diseases / immunology
  • Cytomegalovirus / immunology*
  • Cytomegalovirus Infections / immunology*
  • DNA-Binding Proteins / immunology*
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Humans
  • Immunocompromised Host / immunology
  • Immunoglobulin G / immunology
  • Immunoglobulin M / immunology*
  • Infectious Mononucleosis / immunology
  • Kinetics
  • Radioimmunotherapy
  • Recurrence
  • Rheumatoid Factor / immunology
  • Sensitivity and Specificity
  • Viral Proteins / immunology*

Substances

  • Antibodies, Monoclonal
  • Antibodies, Viral
  • DNA-Binding Proteins
  • ICP36 protein, Cytomegalovirus
  • Immunoglobulin G
  • Immunoglobulin M
  • Viral Proteins
  • Rheumatoid Factor