Integrity of cell-free DNA in maternal plasma extracellular vesicles as a potential biomarker for non-invasive prenatal testing

Large proportions of cell-free DNA (cfDNA) in plasma are localized in extracellular vesicles (EVs), which are secreted from placental cells. This study was conducted to reveal the integrity pattern of cfDNA in maternal plasma EVs (evcfDI) across gestation, and explore if evcfDI could be a potential biomarker in screening for aneuploid fetus in non-invasive prenatal testing (NIPT).

A total of 180 maternal plasma samples were collected during NIPT. Both evcfDNA and fetal evcfDNA (evcffDNA) were measured by quantitative PCR of LINE1 and SRY gene amplicons with different sizes. The evcfDI was calculated as the ratio of long to short fragments.

evcfDI is not affected by gestational age; whereas evcffDI has a mild decreasing trend with increasing gestational age (P = 0.048). evcfDI is significantly and negatively correlated with maternal body mass index (BMI; calculated as weight in kilograms divided by the square of height in meters: ≤18.5, 18.5-25, and ≥25) (P < 0.01) and age (<35 and ≥35 years) (P < 0.01). Mean evcfDI decreases from 2.113 in euploid controls to 0.681 in those with an aneuploid fetus in NIPT (P = 0.003).

Maternal clinical characteristics such as BMI and age could be innovative biomarkers to calibrate evcfDI, which was shown to be a potential indicator of an aneuploid fetus. Analysis of evcfDI based on quantitative PCR could serve as a novel, rapid, and low-cost NIPT strategy, which might facilitate testing at earlier gestations.