dc.date.accessioned | 2023-03-10T17:44:17Z | |
dc.date.available | 2023-03-10T17:44:17Z | |
dc.date.created | 2022-12-21T13:16:04Z | |
dc.date.issued | 2023 | |
dc.identifier.citation | Bitenc, Marie Tune, Benedicte Grebstad Melheim, Maria Atneosen-Åsegg, Monica Lai, Xiaoran Rajar, Polona Solberg, Rønnaug Baumbusch, Lars Oliver . Assessing nuclear versus mitochondrial cell-free DNA (cfDNA) by qRT-PCR and droplet digital PCR using a piglet model of perinatal asphyxia. Molecular Biology Reports. 2022 | |
dc.identifier.uri | http://hdl.handle.net/10852/101219 | |
dc.description.abstract | Abstract
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 (qRT-PCR 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. | |
dc.language | EN | |
dc.publisher | Kluwer Academic/Plenum Publishers | |
dc.rights | Attribution 4.0 International | |
dc.rights.uri | https://creativecommons.org/licenses/by/4.0/ | |
dc.title | Assessing nuclear versus mitochondrial cell-free DNA (cfDNA) by qRT-PCR and droplet digital PCR using a piglet model of perinatal asphyxia | |
dc.title.alternative | ENEngelskEnglishAssessing nuclear versus mitochondrial cell-free DNA (cfDNA) by qRT-PCR and droplet digital PCR using a piglet model of perinatal asphyxia | |
dc.type | Journal article | |
dc.creator.author | Bitenc, Marie | |
dc.creator.author | Tune, Benedicte Grebstad | |
dc.creator.author | Melheim, Maria | |
dc.creator.author | Atneosen-Åsegg, Monica | |
dc.creator.author | Lai, Xiaoran | |
dc.creator.author | Rajar, Polona | |
dc.creator.author | Solberg, Rønnaug | |
dc.creator.author | Baumbusch, Lars Oliver | |
cristin.unitcode | 185,53,46,11 | |
cristin.unitname | Pediatrisk forskningsinstitutt | |
cristin.ispublished | true | |
cristin.fulltext | original | |
cristin.qualitycode | 1 | |
dc.identifier.cristin | 2096335 | |
dc.identifier.bibliographiccitation | info:ofi/fmt:kev:mtx:ctx&ctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.jtitle=Molecular Biology Reports&rft.volume=&rft.spage=&rft.date=2022 | |
dc.identifier.jtitle | Molecular Biology Reports | |
dc.identifier.volume | 50 | |
dc.identifier.issue | 2 | |
dc.identifier.startpage | 1533 | |
dc.identifier.endpage | 1544 | |
dc.identifier.doi | https://doi.org/10.1007/s11033-022-08135-0 | |
dc.type.document | Tidsskriftartikkel | |
dc.type.peerreviewed | Peer reviewed | |
dc.source.issn | 0301-4851 | |
dc.type.version | PublishedVersion | |