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Naslov:Assessing nuclear versus mitochondrial cell-free DNA (cfDNA) by qRT-PCR and droplet digital PCR using a piglet model of perinatal asphyxia
Avtorji:ID Bitenc, Marie (Avtor)
ID Grebstad Tune, Benedicte (Avtor)
ID Melheim, Maria (Avtor)
ID Atneosen‑Åsegg, Monica (Avtor)
ID Rajar, Polona (Avtor), et al.
Datoteke:.pdf PDF - Predstavitvena datoteka, prenos (942,93 KB)
MD5: CA0E79300471BC6E3C2960347E212763
 
URL URL - Izvorni URL, za dostop obiščite https://doi.org/10.1007/s11033-022-08135-0
 
Jezik:Angleški jezik
Tipologija:1.01 - Izvirni znanstveni članek
Organizacija:Logo UKC LJ - Univerzitetni klinični center Ljubljana
Povzetek:Background: Since the discovery more than half a century ago, cell-free DNA (cfDNA) has become an attractive objective in multiple diagnostic, prognostic, and monitoring settings. However, despite the increasing number of cfDNA applications in liquid biopsies, we still lack a comprehensive understanding of the nature of cfDNA including optimal assessment. In the presented study, we continued testing and validation of common techniques for cfDNA extraction and quantification (qRTPCR or droplet digital PCR) of nuclear- and mitochondrial cfDNA (ncfDNA and mtcfDNA) in blood, using a piglet model of perinatal asphyxia to determine potential temporal and quantitative changes at the levels of cfDNA. Methods and Results: Newborn piglets (n=19) were either exposed to hypoxia (n=11) or were part of the sham-operated control group (n=8). Blood samples were collected at baseline (=start) and at the end of hypoxia or at 40–45 min for the sham-operated control group. Applying the qRT-PCR method, ncfDNA concentrations in piglets exposed to hypoxia revealed an increasing trend from 7.1 ng/ml to 9.5 ng/ml for HK2 (hexokinase 2) and from 4.6 ng/ml to 7.9 ng/ml for β-globulin, respectively, whereas the control animals showed a more balanced profile. Furthermore, median levels of mtcfDNA were much higher in comparison to ncfDNA, but without significant differences between intervention versus the control group. Conclusions: Both, qRT-PCR and the droplet digital PCR technique identified overall similar patterns for the concentration changes of cfDNA; but, the more sensitive digital PCR methodology might be required to identify minimal responses.
Ključne besede:cell-free DNA, digital PCR, mitochondrial cfDNA, nuclear cfDNA, perinatal asphyxia, qRT-PCR
Status publikacije:Objavljeno
Verzija publikacije:Objavljena publikacija
Leto izida:2023
Št. strani:str. 1533–1544
Številčenje:Vol. 50, no. 2
PID:20.500.12556/DiRROS-27784 Novo okno
UDK:61:575
ISSN pri članku:1573-4978
DOI:10.1007/s11033-022-08135-0 Novo okno
COBISS.SI-ID:256501507 Novo okno
Opomba:Nasl. z nasl. zaslona; Opis vira z dne 10. 11. 2025;
Datum objave v DiRROS:25.02.2026
Število ogledov:183
Število prenosov:63
Metapodatki:XML DC-XML DC-RDF
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Gradivo je del revije

Naslov:Molecular biology reports
Skrajšan naslov:Mol. biol. rep.
Založnik:Kluwer
ISSN:1573-4978
COBISS.SI-ID:513217817 Novo okno

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Licenca:CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
Povezava:http://creativecommons.org/licenses/by/4.0/deed.sl
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