ArticlePloS one2014
Direct quantification of cell-free, circulating DNA from unpurified plasma.
Article in PloS one, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 77 papers, 2 of them syntheses that pooled it.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
77 citing papers in PubMed, 2 syntheses or guidelines pooled it.
- Circulating free DNA as a diagnostic marker for echinococcosis: a systematic review and meta-analysis.Frontiers in microbiology · 2024Pooled it
- Guidelines for pre-analytical conditions for assessing the methylation of circulating cell-free DNA.Clinical epigenetics · 2021Pooled it
- Validating quantitative PCR assays for cfDNA detection without DNA extraction in exercising SLE patients.Scientific reports · 2021Trial
- Finger-actuated microfluidic device for point-of-care circulating cell-free nucleic acid extraction.Lab on a chip · 2026Article
- PVA sponge method improves detection efficiency of cfDNA in plasma.International journal of clinical oncology · 2026Article
- Preoperative Plasma Cell-Free DNA Integrity Index in Clear Cell Renal Cell Carcinoma: An Exploratory Case-Control Study.Cancers · 2026Article
- Article
- Systematic Review of Methods for Measuring Circulating Cell-Free DNA in Plasma of Healthy Individuals.Diagnostics (Basel, Switzerland) · 2026Review
- Molecular Lung Imaging Following Exposure to Radiation Predicts Long-Term Survival in Rats.International journal of molecular sciences · 2026Article
- Circulating extracellular vesicle DNA (EV-DNA) and cell-free DNA in ovarian cancer: pathological relevance and diagnostic applications.Extracellular vesicles and circulating nucleic acids · 2026Review
- Take charge during treatment: a protocol for a pilot study to evaluate the impact of exercise on cardiovascular biomarkers among black and white breast cancer patients undergoing treatment.Cardio-oncology (London, England) · 2025Article
- Evaluation of automatic cell free DNA extraction metrics using different blood collection tubes.Scientific reports · 2025Article
- A bibliometric analysis of nucleic acid probe and its applications in oncology: towards more precise molecular medicine.Discover oncology · 2025Review
- Methodological influences on circulating cell-free-mitochondrial and nuclear DNA concentrations in response to chronic stress.Molecular biology reports · 2025Article
- Circulating Cell-Free DNA in Metabolic Diseases.Journal of the Endocrine Society · 2025Review
- A Single Microfluidic Device Approach to Direct Isolation, Purification, and Amplification of cfDNA from Undiluted Plasma.Sensors and actuators. B, Chemical · 2025Article
- Stimulation of mouse hematopoietic stem cells by angiogenin and DNA preparations.Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologica · 2024Article
- Article
- Detecting liquid remnants of solid tumors treated with curative intent: Circulating tumor DNA as a biomarker of minimal residual disease (Review).Oncology reports · 2023Review
- Genome-wide tiled detection of circulating Mycobacterium tuberculosis cell-free DNA using Cas13.Nature communications · 2023Article
17 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
10 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Cell-free DNA (cfDNA) in body tissues or fluids is extensively investigated in clinical medicine and other research fields. In this article we provide a direct quantitative real-time PCR (qPCR) as a sensitive tool for the measurement of cfDNA from plasma without previous DNA extraction, which is known to be accompanied by a reduction of DNA yield. The primer sets were designed to amplify a 90 and 222 bp multi-locus L1PA2 sequence. In the first module, cfDNA concentrations in unpurified plasma were compared to cfDNA concentrations in the eluate and the flow-through of the QIAamp DNA Blood Mini Kit and in the eluate of a phenol-chloroform isoamyl (PCI) based DNA extraction, to elucidate the DNA losses during extraction. The analyses revealed 2.79-fold higher cfDNA concentrations in unpurified plasma compared to the eluate of the QIAamp DNA Blood Mini Kit, while 36.7% of the total cfDNA were found in the flow-through. The PCI procedure only performed well on samples with high cfDNA concentrations, showing 87.4% of the concentrations measured in plasma. The DNA integrity strongly depended on the sample treatment. Further qualitative analyses indicated differing fractions of cfDNA fragment lengths in the eluate of both extraction methods. In the second module, cfDNA concentrations in the plasma of 74 coronary heart disease patients were compared to cfDNA concentrations of 74 healthy controls, using the direct L1PA2 qPCR for cfDNA quantification. The patient collective showed significantly higher cfDNA levels (mean (SD) 20.1 (23.8) ng/ml; range 5.1-183.0 ng/ml) compared to the healthy controls (9.7 (4.2) ng/ml; range 1.6-23.7 ng/ml). With our direct qPCR, we recommend a simple, economic and sensitive procedure for the quantification of cfDNA concentrations from plasma that might find broad applicability, if cfDNA became an established marker in the assessment of pathophysiological conditions.
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