ABSTRACT: Indirect testing for elevated sera levels of antisperm antibody assumes that all relevant antigens are present on the target sperm utilized. In the present study, the heterogeneity of reactivity of positive sera with sperm from different donors was addressed. When 68 sera that previously tested positive by either sperm immobilization test (SIT) or ELISA or both were tested for antisperm antibody levels by a quantitative ELISA using nine different sperm donors, the frequency of positive reactions was 51% for men and 81% for women. A 50% correlation of SIT- and ELISA-positive results could be improved to 85% using the same sperm specimen. This would improve the overall correlation of functional and ELISA test results from 95% to 99% in the infertile population studied. The data suggest that individual sperm may vary in their antigenic expression and that comparison of methods between laboratories could be improved if equivalent target sperm were used.