The present study was conducted to estimate the copy number of mitochondrial COXI gene in the sperm cells from Iraqi asthenozoospermic patients by using q PCR. Cases were established of Asthenozoospermia from Kamal AL.Samraee Hospital for Infertility and In Vitro Fertilization in Baghdad-Iraq. The relative quantification of mtDNA copy number was performed for 28 sperm extracts (28 fractions 80 % layers and 28 fractions 40% layers) obtained from 28ejaculates, 16 samples were from asthenozoospermic patients and 12 samples were from control subjects. The content of mtDNA per haploid mitochondrial genome in sperm cell was significantly different (p≤ 0.05) between non- progressive motile sperm cells and mtDNA content corresponded to the progressive sperm (119.9 ± 26.89 and 58.99 ± 22.68, respectively). Increasing of relative mtDNA copy number of defective sperm fraction and normospermic fractions were 74.2-271.3 and 1.9-92.8/copy per cell, respectively. No significant difference between progressive motile sperms of infertile subjects and progressive motile sperms of control subjects that were isolated from 80% percoll layer as well as to non progressive motile sperms of both groups that were separated from 40% Percoll fraction. There is a highly significant difference (p≤0.05) between infertile subject and control subject on average 128.4 ± 14.76 and 60.90 ± 6.83 copy per cell.