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Detection of antisperm antibodies in seminal plasma by flow cytometry: comparison with the indirect immunobead
S C Nicholson1, J N Robinson, I L Sargent
1Nuffield Department of Obstetrics and Gynaecology, John Radcliffe Maternity Hospital, Oxford, United Kingdom.
Objective:
To compare flow cytometry with the established indirect immunobead binding test (IBT) for the detection of antisperm antibodies in seminal plasma.
Design:
A prospective, comparative study.
Setting:
University-based andrology unit.
Patient(S):
One hundred and fifty-eight men with suspected male factor subfertility.
Intervention(S):
Seminal plasma samples were incubated with antisperm antibody-negative donor sperm. Surface-bound antibody was detected with fluorescence-labeled antihuman antibody in the flow cytometry assay or with immunobead-labeled antihuman antibody in the IBT.
Main Outcome Measure(S):
The percentage of sperm that tested positive for surface-bound antibody was determined in the two assays. Seminal plasma was antisperm antibody-positive when > or = 20% of the sperm were antibody-bound, and clinically significant levels were present when > or = 50% of the sperm were antibody-bound.
Result(S):
Of 71 samples that were negative by the IMT, 66 (93%) also were negative by flow cytometry. Of 63 samples that had > or = 50% immunobead binding, 55 had equivalent results by flow cytometry. Overall statistical analysis showed a good correlation between the two assays.
Conclusion(S):
There is a good correlation between the indirect IBT and indirect flow cytometry for the detection of antisperm antibodies in seminal plasma.