Background : Prenatal paternity test is mostly performed by using Amniocentesis or Chorionic Villus Sampling (CVS) methods. However, these methods require invasive procedures, which are potentially harmful for both the mother and the fetus. Currently, the invention of of Cell-Free Fetal DNA (cffDNA) has offered the opportunity of performing prenatal paternity test non-invasively. Materials and Methods : This study is a cross sectional descriptive study to detect cell free fetal DNA at 13 STR loci and at amelogenin gene to evaluate fetus gender, which will be compared to the baby gender afterbirth. Healthy third semester pregnant women were included as participants. Inform consent for both the mother and the biological father has been provided. Result : Four participants has been evaluated. In this study, in all participants, we found the presence of cffDNA in almost all of the STR loci. Some loci cannot be detected due to the small amount of cffDNA in the loci. All fetus genders detected by cffDNA in the amelogenin gene macthed the gender of the four babies afterbirth. Conclusion : The use of Cell-Free Fetal DNA (cffDNA) is a potential non-invasive methods in prenatal paternity test. Additionally, the ability of the method to evaluate fetus gender has been suggested.