Donor-specific antibody detection: comparison of single antigen assay and Luminex crossmatches.
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.
Caro-Oleas
J L
JL
Servicio de Inmunología, Instituto de Biomedicina (IBIS), Hospital Universitario Virgen del Rocío, Universidad de Sevilla, Spain. joseluiscaro@hotmail.com
González-Escribano
M F
MF
Toro-Llamas
S
S
Acevedo
M J
MJ
Martinez-Bravo
M J
MJ
Aguilera
I
I
Núñez-Roldán
A
A
D000920:Antibody-Dependent Cell Cytotoxicity; D003165:Complement System Proteins / Q000378:metabolism; D003601:Cytotoxicity Tests, Immunologic; D005260:Female; D015234:HLA-A Antigens / Q000276:immunology; D015235:HLA-B Antigens / Q000276:immunology; D006683:HLA-DQ Antigens / Q000276:immunology; D006684:HLA-DR Antigens / Q000276:immunology; D006650:Histocompatibility Testing / Q000379:methods* / Q000706:statistics & numerical data; D006801:Humans; D007518:Isoantibodies / Q000097:blood*; D011247:Pregnancy; D012680:Sensitivity and Specificity; D014019:Tissue Donors; D014181:Transplantation Immunology
D015234:HLA-A Antigens; D015235:HLA-B Antigens; D006683:HLA-DQ Antigens; D006684:HLA-DR Antigens; D007518:Isoantibodies; D003165:Complement System Proteins