Flow cytometry provides rapid and highly accurate detection of antisperm antibodies.
OBJECTIVE
Immunobead testing (IBT), the current standard for antisperm antibody detection, is time consuming and somewhat subjective. To overcome these limitations and maintain accuracy, we studied an immunofluorescent assay using flow cytometry.


METHODS
A validation study comparing flow cytometry to IBT in the detection of serum antisperm antibodies.


METHODS
Flow cytometry laboratory.


METHODS
Sera from 37 men after vasectomy (test) and sera from 35 fertile men (control).


METHODS
Test serum with and without immunoglobulin (Ig)G, IgA, and IgM antisperm antibodies as defined by IBT were analyzed by flow cytometry. Sensitivity and specificity of flow cytometry was calculated by defining the IBT as the true result.


RESULTS
Flow cytometry identified 22 of 22 sera that were IgG positive (100% sensitivity), 12 of 14 sera that were IgA positive (86% sensitivity), and 4 of 4 sera that were IgM positive (100% sensitivity). Overall, 22 of 37 men were positive for antisperm antibodies. The flow cytometry correctly identified 71 of 71 negative sera (100% specificity). Fluorescence intensity values from the 37 study patients significantly correlated with immunobead binding to the head region and to the entire (more than one) region.


CONCLUSIONS
Detection of IgG, IgA, and IgM antisperm antibodies by flow cytometry is highly sensitive and specific. In addition, flow cytometry is able to assess thousands of sperm rapidly and accurately, reducing sampling error and technical time.
Ke
R W
RW
University of Tennessee, Memphis.
Dockter
M E
ME
Majumdar
G
G
Buster
J E
JE
Carson
S A
SA
D000906:Antibodies / Q000032:analysis*; D005069:Evaluation Studies as Topic; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D007070:Immunoglobulin A / Q000032:analysis; D007074:Immunoglobulin G / Q000032:analysis; D007075:Immunoglobulin M / Q000032:analysis; D007158:Immunologic Techniques; D008297:Male; D008863:Microspheres; D012680:Sensitivity and Specificity; D013094:Spermatozoa / Q000276:immunology*; D013997:Time Factors
D000906:Antibodies; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007075:Immunoglobulin M