Ginekologia i Poloznictwo
ISSN 1896-3315 e-ISSN 1898-0759

Laser assisted ALU-qPCR to evaluate the association between cell-free DNA in human follicular fluid with oocyte and embryo quality in assisted reproductive techniques


Abstract

Author(s): Rabab M. Sorour, Mohamed A. El Noury, Osama Fekry, Sondos Salem, Tamer Taha and Karim Abdelsalam*

Background: Cell-free DNA (cfDNA) has the potential to provide non-invasive diagnostic assays and practical biomarkers for predicting the outcome of assisted reproductive technology (ART) procedures. Aim of the study: To investigate the association between cf-DNA concentration in individual follicular fluid and oocyte maturity and embryo quality in patients undergoing Intra Cytoplasmic Sperm injection (ICSI).

Patients and methods: This prospective observational clinical study was done at Ain Shams University, A maternity hospital (IVF Unit), and a private ICSI center from May 2019 to June 2020. Our study used the ALU-QPCR technique to accurately measure the cell-free DNA (cfDNA) level in 35 follicular fluid samples from 15 patients undergoing ICSI.

Results: Our results displayed the range of cfDNA from 0.24 to 3.89 (ng/ml), with a mean of 2.06 ± 0.73. The DNA integrity (Q247/Q115) ranged from 0.09 to 0.79, with a mean of 0.44 ± 0.18. Our findings reveal a statistically significant association between the levels of cell-free DNA and patient oocytes to yield an Embryo, top grade Embryos, Fragmentation rate of the embryo, and number of blastomeres (P-values; p=0.037, p<0.001, p<0.001, and p=0.019, respectively ). A statistically negative correlation between cfDNA (ng/ml) and follicle size in mm (p-value < 0.05). However, no significant correlation was observed for the other parameters (basic hormonal profile, AFC number of oocytes, and the number of fertilized oocytes), with a p-value > 0.05. The study found that the levels of cfDNA and 17β-estradiol had a non-significant negative correlation (p-value > 0.05). Our study found that the level of cfDNA in follicular fluid is strongly associated with the quality of embryos on Day 3.

Conclusion: High levels of cfDNA in follicular fluid may indicate poor follicular milieu. Measuring cfDNA can evaluate oocyte and embryo competence. It can be a non-invasive test to assess embryo quality along with morphological criteria.

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