An improved indirect enzyme-linked immunosorbent assay (ELISA) for the detection of antisperm antibodies

An indirect enzyme-linked immunosorbent assay (ELISA) was used for the detection of antisperm antibodies which provides a sensitivity and ease of quantitation that is not available with conventional bioassay systems. A standardized protocol was developed in which washed whole sperm were coated onto polystyrene microelisa plates at a density of 1 X 10(5) sperm per well using a commercially available spray fixative. Urease was employed in the enzyme-antiimmunoglobulin conjugate to minimize nonspecific background reactions. Diluted positive and negative human sera were incubated at 37 degrees C and the results were read on an ELISA auto reader. Using mouse antihuman sperm antisera and sera from selected infertile patients, it was found that the ELISA method was significantly more sensitive than the sperm microimmobilization test and the microtray agglutination test. The results also confirmed that the ELISA detected a different spectrum of sperm antibodies compared with the other two techniques.