OBJECTIVE: To assess the use of flow cytometry (FCM) analysis of living spermatozoa subjected to indirect immunofluorescence staining with the mouse anti-human immunoglobulin (Ig)G monoclonal antibodies (FCM test) as an objective method for the detection of antisperm antibodies in semen and serum. DESIGN: The comparative studies were conducted with the direct and indirect mixed antiglobulin reaction (MAR) and FCM test performed by FCM analyzer FACSCAN (Becton Dickinson Immunocytometry Systems, Mountain View, CA). SETTING: Clinical infertile service. PATIENTS: Seventy-six semen and 20 sera samples of infertile patients were analyzed. RESULTS: For IgG in semen there was a highly significant correlation between the direct MAR and FCM test. Six of 12 IgG-positive samples revealed also IgA, and just one sample, which showed negative MAR, was positive for IgM in FCM test. Flow cytometry test and indirect MAR showed also a highly significant correlation of the results of IgG detection in sera. CONCLUSIONS: Flow cytometry test may be used to verify that IgG antisperm antibodies are on the surface of living spermatozoa either retrieved directly from an ejaculate or after exposure to serum and to determine proportion of antisperm antibodies-positive spermatozoa, titer, and quantity of antibodies bound to the cell surface.
OBJECTIVE: To assess the use of flow cytometry (FCM) analysis of living spermatozoa subjected to indirect immunofluorescence staining with the mouse anti-human immunoglobulin (Ig)G monoclonal antibodies (FCM test) as an objective method for the detection of antisperm antibodies in semen and serum. DESIGN: The comparative studies were conducted with the direct and indirect mixed antiglobulin reaction (MAR) and FCM test performed by FCM analyzer FACSCAN (Becton Dickinson Immunocytometry Systems, Mountain View, CA). SETTING: Clinical infertile service. PATIENTS: Seventy-six semen and 20 sera samples of infertilepatients were analyzed. RESULTS: For IgG in semen there was a highly significant correlation between the direct MAR and FCM test. Six of 12 IgG-positive samples revealed also IgA, and just one sample, which showed negative MAR, was positive for IgM in FCM test. Flow cytometry test and indirect MAR showed also a highly significant correlation of the results of IgG detection in sera. CONCLUSIONS: Flow cytometry test may be used to verify that IgG antisperm antibodies are on the surface of living spermatozoa either retrieved directly from an ejaculate or after exposure to serum and to determine proportion of antisperm antibodies-positive spermatozoa, titer, and quantity of antibodies bound to the cell surface.