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Published on: November 27, 2017
Amniotic fluid cell-free DNA in preterm prelabor rupture of membranes
Marian Kacerovsky1,2, Barbora Vlkova3, Ivana Musilova1
1Department of Obstetrics and Gynecology, Charles University, Faculty of Medicine Hradec Kralove, University Hospital Hradec Kralove, Hradec Kralove, Czech Republic.
Introduction:
We evaluated the levels of cell-free nuclear DNA (nDNA) and cell-free mitochondrial DNA (mtDNA) in the amniotic fluid supernatant from pregnancies complicated by preterm prelabor rupture of membranes (PPROM) based on evidence of microbial invasion of the amniotic cavity (MIAC) and/or intra-amniotic inflammation (IAI).
Material And Methods:
A total of 155 women with PPROM were included in this study. Amniotic fluid samples were obtained by transabdominal amniocentesis. The levels of cell-free nDNA and mtDNA in the amniotic fluid supernatant were assessed and quantified by real-time polymerase chain reaction.
Results:
The levels of cell-free nDNA and mtDNA were higher in women with MIAC and IAI than in women without these conditions (nDNA: with MIAC: median 3.9 × 104 genome equivalent [GE]/mL vs without MIAC: median 1.2 × 104 GE/mL, with IAI: median: 5.3 × 104 GE/mL vs without IAI: median 1.2 × 104 GE/mL; mtDNA: with MIAC: median 9.2 × 105 GE/mL vs without MIAC: median 2.5 × 105 GE/mL, with IAI: median 1.1 × 106 GE/mL vs without IAI: median 2.5 × 105 ; all P values ≤ 0.01). Women with the microbial-associated IAI showed the highest levels of cell-free nDNA and mtDNA.
Conclusions:
Cell-free nDNA and mtDNA are constituents of the amniotic fluid supernatant from PPROM pregnancies. Both cell-free nDNA and mtDNA are involved in the intra-amniotic inflammatory response in women with PPROM.
Insights
Cell-free nuclear DNA (nDNA) and mitochondrial DNA (mtDNA) are elevated in amniotic fluid during preterm prelabor rupture of membranes (PPROM) with microbial invasion or inflammation. These DNA fragments play a role in the inflammatory response in PPROM pregnancies.
Area of Science:
- Obstetrics and Gynecology
- Maternal-Fetal Medicine
- Molecular Diagnostics
Background:
- Preterm prelabor rupture of membranes (PPROM) is a major cause of preterm birth.
- Microbial invasion of the amniotic cavity (MIAC) and intra-amniotic inflammation (IAI) are significant complications of PPROM.
- Cell-free DNA (cfDNA) in amniotic fluid may serve as a biomarker for these complications.
Purpose of the Study:
- To evaluate cell-free nuclear DNA (nDNA) and cell-free mitochondrial DNA (mtDNA) levels in amniotic fluid supernatant from PPROM pregnancies.
- To correlate cfDNA levels with evidence of MIAC and/or IAI.
- To understand the role of cfDNA in the intra-amniotic inflammatory response.
Main Methods:
- Amniotic fluid samples were collected from 155 women with PPROM via transabdominal amniocentesis.
- Real-time polymerase chain reaction (PCR) was used to quantify cell-free nDNA and mtDNA levels.
- Samples were analyzed for the presence of MIAC and IAI.
Main Results:
- Significantly higher levels of cell-free nDNA and mtDNA were observed in women with MIAC and IAI compared to those without.
- Specific median concentrations: nDNA with MIAC (3.9 × 10⁴ GE/mL) vs. without (1.2 × 10⁴ GE/mL); mtDNA with MIAC (9.2 × 10⁵ GE/mL) vs. without (2.5 × 10⁵ GE/mL).
- Women with microbial-associated IAI exhibited the highest cfDNA levels.
Conclusions:
- Cell-free nDNA and mtDNA are present in the amniotic fluid supernatant of PPROM pregnancies.
- Both cfDNA types are implicated in the intra-amniotic inflammatory response associated with PPROM.
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