A prospective study of non-invasive preimplantation genetic testing for Rh status, sex prediction and mtDNA copy number analysis using ddPCR on spent culture media
EXPERIMENTAL AND THERAPEUTIC MEDICINE, cilt.32, sa.3, 2026 (SCI-Expanded)
- Yayın Türü: Makale / Tam Makale
- Cilt numarası: 32 Sayı: 3
- Basım Tarihi: 2026
- Doi Numarası: 10.3892/etm.2026.13245
- Dergi Adı: EXPERIMENTAL AND THERAPEUTIC MEDICINE
- Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), BIOSIS, EMBASE, Academic Search Ultimate (EBSCO), Biomedical Reference Collection: Corporate Edition (EBSCO), Health Research Premium Collection (ProQuest)
- Açık Arşiv Koleksiyonu: AVESİS Açık Erişim Koleksiyonu
- İstanbul Üniversitesi Adresli: Evet
Özet
Presence of embryo-derived cell-free DNA (cfDNA) in spent culture medium (SCM) represents a promising non-invasive alternative to the conventional preimplantation genetic testing (PGT). The present study evaluated the feasibility of using cfDNA isolated from SCM to determine Rhesus (Rh) status through Rh blood group D antigen (RHD) gene analysis, predicting sex-linked disorders using sex determining region Y (SRY) gene detection and assessing the association between mitochondrial cfDNA (cf-mtDNA) levels and maternal age using mitochondrially encoded cytochrome b (MT-CYB) gene quantification. SCM samples collected during the IVF process were subjected to cfDNA isolation and target regions including RHD exon 7 and exon 10, SRY and MT-CYB were analyzed using droplet digital PCR. RHD gene amplification was detected in 6 of 15 samples for exon 7 and in 11 samples for exon 10, with partial concordance between exons. SRY amplification was observed in 16 of 31 samples and was largely consistent with clinical sex data. MT-CYB was detected in 19 of 31 samples, with higher cf-mtDNA/nuclear cfDNA ratios observed in women >40 years of age. A moderate positive correlation trend was noted between maternal age and MT-CYB levels (r=0.46; P=0.354). These findings demonstrate the potential applicability of SCM-derived cfDNA for non-invasive Rh and sex determination without compromising embryo integrity, while mtDNA quantification may provide additional insights into embryo quality. Overall, SCM-based cfDNA analysis exhibited promise as a complementary tool for embryo selection in IVF workflows.