• Antisperm antibody;
  • detection;
  • membrane domains;
  • ELISA;
  • localization

ABSTRACT: Human sera were identified as positive or negative for sperm-reactive antibodies in a solid-phase enzyme-linked immunosorbent assay (ELI-SA). Of these, 28 positive and 22 negative sera were blind-coded and used as first antibody to compare three immunoassays, a modified liquid-phase indirect immunobead assay (IRA); a liquid-phase indirect immunofluorescence assay (IFA); and a solid-phase indirect immunogold assay (IGA). These three immunoassays perform both as sperm-reactive antibody detection assays and as sperm-associated antigen localization assays. As antibody detection assays, the IBA, IFA, and IGA gave 37, 27, and 28 positives and 13, 23, and 22 negatives, respectively. The usefulness of the IBA as an antigen localization assay was limited by the size of the marker, while the smaller IFA and IGA markers enabled increased resolution of binding patterns of sperm-reactive antibodies to surface-associated sperm antigens. Although the antigen-antibody binding patterns were almost identical for IFA and IGA, suggesting the same sperm-associated antigens were detected by both assays, the IGA reaction product was stable, higher in resolution, and visible by light microscopy.