Assessing loss of imprint methylation in sperm from subfertile men using novel methylation polymerase chain reaction Luminex analysis.

摘要

OBJECTIVE: To assess the clinical value of bisulfite polymerase chain reaction Luminex (BPL), an automated, high-throughput procedure for the detection of alterations in DNA methylation. DESIGN: Experimental prospective study. SETTING: University research laboratory and private in vitro fertilization (IVF) clinic. PATIENT(S): A total of 337 men, 61 with severe oligozoospermia, 67 with moderate oligozoospermia, and 209 with microscopically normozoospermia. INTERVENTION(S): The ejaculated sperm samples after the routine semen analysis with patients' consent. MAIN OUTCOME MEASURE(S): Examination of the methylation patterns of eight imprinted loci in sperm DNA, and confirmation with combined bisulfite PCR restriction analysis (COBRA). RESULT(S): A total of 47 cases (13.9%) showed abnormal methylation at one or more imprinted loci (18 paternal, 18 maternal, and 11 cases with alterations of both maternal and paternal imprints). CONCLUSION(S): The relative ease of the BPL method provides a practical method within a clinical setting to reduce the likelihood of abnormal samples being used in assisted reproduction treatments.