POLYMERASE CHAIN-REACTION ENZYME-LINKED-IMMUNOSORBENT-ASSAY DETECTIONOF MYCOPLASMA CONSENSUS GENE IN SPERM WITH LOW OOCYTE PENETRATION CAPACITY

Objective: To determine the prevalence of mycoplasmas in washed sperm and to compare the penetration of zona-free hamster oocytes by sperm w ith and without mycoplasmas. Design: Prospective comparative study. Se tting: Clinical and academic research environment. Patient(s): Semen f rom 34 male patients. Intervention(s): Specimens were divided, Percoll washed, and scanned for differences in kinematic parameters. Sperm DN A was extracted and assayed for mycoplasma DNA using the polymerase ch ain reaction-ELISA method targeting the consensus gene of 15 mycoplasm a species. Remaining sperm were processed by centrifuge, Percoll, or T EST (TES and Tris) Yolk Buffer (TYB) and assessed for penetration capa city. Main Outcome Measure(s): Detection of mycoplasma DNA. Result(s): Mycoplasma DNA was detected in 29.4% of the Percoll-washed sperm. The penetration of oocytes by mycoplasma-positive sperm (59.5% +/- 17.3%; mean +/- SEM) was less than mycoplasma-negative sperm (86.8% +/- 5.4% ) in the TYB-processed group. Conclusion(s): Mycoplasma DNA is demonst rated in almost a third of the Percoll-washed sperm. Because there wer e no other cell types except sperm, the results suggest that the mycop lasmas were either internalized or attached to the membranes. The redu ced penetration by mycoplasma-positive sperm after 48-hour TYB suggest mycoplasmas required time to affect sperm function. Similarities betw een hypo-osmotic swelling and between kinematic parameters suggest tha t the mechanism does not involve differences in membrane integrity and in motility patterns.