Comparison of Cell-Free Fetal DNA Plasma Content Used to Sex Determination Between Three Trimesters of Pregnancy in Torkaman Pregnant Mare.

This investigation aimed to compare the cell-free fetal DNA (cffDNA) plasma present in three trimesters of pregnancy in Torkaman pregnant mare. Peripheral blood samples of 32 pregnant mares in three trimesters of pregnancy were collected in tubes containing ethylenediaminetetraacetic acid at three time points. Circulating cffDNA was extracted from 3 mL of maternal plasma. Using outer and inner primers, a conventional polymerase chain reaction was performed for the sex-determining region Y (SRY) gene present in the Y chromosome. Of the total 32 Torkaman pregnant mares, 24 were carrying male fetuses and eight were carrying female fetuses. In total, the accuracy of the test was 48.75%, 68.75%, and 75% in the first, second, and third trimesters of pregnancy, respectively. The sensitivities were 25%, 58.32%, and 66.66%, respectively, whereas their specificities were 100% in all trimesters. In conclusion, the SRY gene can permit the detection of equine fetal sex with good accuracy through cffDNA analysis in maternal plasma just in the third trimester of pregnancy, although specificity in all duration of pregnancy was 100%. • cffDNA, Eca.YH12, and TKY270 cannot be used in 1st and 2nd trimesters of pregnancy. • cffDNA, Eca.YH12, and TKY270 is better for female sex determination • cffDNA, Eca.YH12, and TKY270 cannot be used in 3rd trimester of pregnancy. • cffDNA, Eca.YH12, and TKY270 was detected with 100% specificity in all trimesters. [ABSTRACT FROM AUTHOR]