Non-Invasive Fetal Sex Determination Using SRY Specific Primers and Sybrgreen Real Time PCR

Abstract

Presence of fetal cells and circulating free fetal DNA and RNA in maternal circulation represents the basic concept in developement of non-invasive prenatal diagnostic methods based on molecular biology and genetics. We introduced new methods for free fetal DNA isolation and detection in maternal circulation via DNA isolation from maternal plasma using real-time PCR SYBRGreen targeting and newly designed primers focused in SRY sequence. We determined gender in 46 singleton pregnancies, 22 boys and 24 girls and assessed the analytical and clinical validity. We reached 95.45% sensitivity and 95.83% specificity. We suggest improvements in molecular-biological procedures in the discussion, which could be used in studies of clinical utility of non-invasive prenatal diagnosis (NIPD) in decreasing amount of invasive procedures unnecessarily performed.