Donor-specific antibody detection: comparison of single antigen assay and Luminex crossmatches

Tissue Antigens. 2010 Nov;76(5):398-403. doi: 10.1111/j.1399-0039.2010.01531.x. Epub 2010 Aug 19.

Abstract

Luminex bead-based assays are routinely used in the study of anti-human leukocyte antigen (HLA) donor-specific antibodies (DSA). Single antigen (SA) assays use beads coated with recombinant antigens whereas Luminex crossmatch (Xm-DSA) tests consist of beads coated with isolated donor-specific HLA molecules. The aim of this study was to compare these techniques used to detect DSA. A total of 24 sera recognizing different HLA class I (seven anti-HLA-A and seven anti-HLA-B) as well as class II (seven anti-HLA-DR and three anti-HLA DQ) specificities by complement dependent cytotoxicity were included in the study. These sera were used undiluted and in serial dilutions to perform both class I and II SA and Xm-DSA assays. In the case of Xm-DSA the same serum was checked with different lysates. A total of 42 lysates were used to perform a total of 61 crossmatches: 42 to detect anti-class I and 19 to detect anti-class II antibodies. The maximum positive dilution was higher for SA in 76% of the class I and in 90% of the class II crossmatches. Those cases with a higher sensitivity of the Xm-DSA could not be explained by a larger number of antigen targets.

Publication types

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

MeSH terms

  • Antibody-Dependent Cell Cytotoxicity
  • Complement System Proteins / metabolism
  • Cytotoxicity Tests, Immunologic
  • Female
  • HLA-A Antigens / immunology
  • HLA-B Antigens / immunology
  • HLA-DQ Antigens / immunology
  • HLA-DR Antigens / immunology
  • Histocompatibility Testing / methods*
  • Histocompatibility Testing / statistics & numerical data
  • Humans
  • Isoantibodies / blood*
  • Pregnancy
  • Sensitivity and Specificity
  • Tissue Donors*
  • Transplantation Immunology

Substances

  • HLA-A Antigens
  • HLA-B Antigens
  • HLA-DQ Antigens
  • HLA-DR Antigens
  • Isoantibodies
  • Complement System Proteins