A Multi-Laboratory Evaluation of an Enzyme-Linked Immunoassay Quantitating Human Antibodies to Streptococcus Pneumoniae Polysaccharides

Immunol Invest. 2001 Aug;30(3):191-207. doi: 10.1081/imm-100105064.

Abstract

An enzyme-linked immunoassay (EIA) is described and evaluated which quantitates human antibodies to serotype specific S. pneumoniae polysaccharide (PnPs) in human sera. Based on the observations previously described by Koskela (1), native PnPs are used as coating antigens and sera are absorbed with a soluble pneumococcal absorbant material containing C-polysaccharide (CPs) to ensure measurement of serotype specific anti-PnPs antibodies. The robustness of this method was evaluated by ten laboratories using the same reagents, protocol, and five human serum samples. Reproducible antibody values were obtained for IgM, IgG, and IgA antibodies to five different PnPs serotypes, 3, 6B, 14, 19F, and 23F. The overall mean percent coefficients of variation in this interlaboratory study for all five selotype specific anti-PnPs determinations with the five coded sera were 30% for IgG, 3/% for IgM, and 36% for IgA. This assay can be standardized for quantitation of serotype specific anti-PnPs antibodies, allowing comparison of antibody values in vaccine trials evaluating pneumococcal vaccines.

Publication types

  • Evaluation Study

MeSH terms

  • Enzyme-Linked Immunosorbent Assay / methods*
  • Evaluation Studies as Topic
  • Humans
  • Immune Sera / classification
  • Immune Sera / immunology
  • Immunoglobulin A / analysis
  • Immunoglobulin G / analysis
  • Immunoglobulin Isotypes / classification
  • Immunoglobulin M / analysis
  • Laboratories / statistics & numerical data
  • Polysaccharides, Bacterial / immunology*
  • Reproducibility of Results
  • Serotyping / methods*
  • Streptococcus pneumoniae / immunology*

Substances

  • Immune Sera
  • Immunoglobulin A
  • Immunoglobulin G
  • Immunoglobulin Isotypes
  • Immunoglobulin M
  • Polysaccharides, Bacterial