In previous years, identification of fetal cells in maternal blood circulation 
has caused a new revolution in non-invasive method of prenatal diagnosis. Low 
number of fetal cells in maternal blood and long-term survival after pregnancy 
limited the use of fetal cells in diagnostic and clinical applications. With the 
discovery of cell-free fetal DNA (cffDNA) in plasma of pregnant women, access to 
genetic material of the fetus had become possible to determine early gender of a 
fetus in pregnancies at the risk of X-linked genetic conditions instead of 
applying invasive methods. Therefore in this study, the probability of detecting 
sequences on the Y chromosome in pregnant women has been evaluated to identify 
the gender of fetuses. Peripheral blood samples were obtained from 80 pregnant 
women at 6(th) to 10(th) weeks of gestation and then the fetal DNA was extracted 
from the plasma. Nested PCR was applied to detect the sequences of single copy 
SRY gene and multi copy DYS14 & DAZ genes on the Y chromosome of the male 
fetuses. At the end, all the obtained results were compared with the actual 
gender of the newborns. In 40 out of 42 born baby boys, the relevant gene 
sequences were identified and 95.2% sensitivity was obtained. Non-invasive early 
determination of fetal gender using cffDNA could be employed as a pre-test in 
the shortest possible time and with a high reliability to avoid applying 
invasive methods in cases where a fetus is at the risk of genetic diseases.