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AIM AND BACKGROUND: The aim of this study was to construct the differential expression profile of sperm miRNA between HIV/AIDS patients and healthy individuals.MATERIALS AND METHODS: The semen samples were obtained from 34 patients infected with HIV/AIDS and 20 age-matched controls.Total RNA were extracted to construct the differential expression profile of miRNA by using gene chip technology and identified by using real-time fluorescent quantitative PCR (qPCR).The differential expression significant miRNAs are selected and compared with both viral and human miRNAs in miRBase database to determine their classification.RESULTS: High mobility and low mobility sperm were successfully separated without somatic cell contamination.The results of miRNA differential expression profile data analysis showed that there were 77 miRNAs which were up-regulated and hiv1-miR-H1, with a fold 51.262568 difference in expression, was the most different, while there were 81 miRNAs which were down-regulated, and has has-miR-1299, with a fold 0.006907 difference in expression was the most distinguishing.Eleven candidate miRNAs were selected for verification based on the finding of up or down regulation in the spermatozoa of HIV patients: hsa-miR-155-5p and hsa-miR-107 were increased in HIV patients, and hsa-miR-1299, has-miR-513a-3p, has-miR-148a-3p, has-miR-106a-5p, has-miR-221-5p were decreased which they were consonant with the result of gene chip assay.