The Potential Usefulness of Free Fetal DNA for Prenatal Fetal Gender Determination in Blood Plasma of Pregnant Goat

To date, sex determination technique in domestic animal has economic benefits. Recently, circulating free DNA (cfDNA) highlighted as suitable candidate for non-invasive sex determination in livestock and endangered wild life animals. Based on this motivation, the objective of this report was the potential usefulness of free fetal DNA in maternal blood for prenatal fetal gender determination in pregnant goats.

 In this respect, a total of 21 animals, including 17 pregnant females with a gestation period of six weeks, unborn goats and 2 males (negative controls) were selected. During the running experiment, estrous synchronization  was performed by intravaginal CIDR. The animals not returned to estrus was considered as an indicator of success in pregnancy. Also in the last months, ultrasound method was used to confirm pregnancy. In the next phase, cffDNA was extracted from the blood plasma of female goats using the available common experimental protocol. The primers in this experiment were designed based on AME gene located on X and Y chromosomes. The polymerase chain reaction was optimized based on available standard methods.

The observation addressed existence of two fragment in the male embryo (171 bp AMELX gene band and 111 bp AMELY gene band) and one fragment in the female embryo (171 bp AMELX gene band). Here, most of findings was in line of similar pervious literatures and sex determination based on AME gene in case of sensitivity, specificity and accuracy indices indicated 0.83, 0.93 and 0.93 respectively.

On this basis, the results opened a new horizon of the potential usefulness of free fetal DNA in maternal blood for prenatal fetal gender determination in pregnant goats.