Purification and quantification of circulating cell-free DNA from body fluids with DANAGENE Circulating System applied to Liquid Biopsy

: Cell-free DNA (cfDNA) is present in numerous body fluids in addition to initiates generally from blood cells. It is undoubtedly the utmost promising tool among all components of liquid biopsy. Liquid biopsy is a specialized method investigating the nonsolid biological tissue by revealing of circulating cells, cell free DNA etc. that enter body fluids. Since, cancer cells disengage from compact tumors circulate in peripheral blood, evaluating blood of cancer patients holds the opportunities for capture and molecular level analysis of various tumor-derived constituents. Cell free DNA samples can deliver a significant perceptions into oncology, for instance tumor heterogeneity, instantaneous tumor development, response to therapy and treatment, comprising immunotherapy and mechanisms of cancer metastasis. Malignant growth at any phase can outhouse tumor cells in addition to fragments of neoplasticity causing DNA into circulatory system giving noble sign of mutation in the tumor at sampling time. Liquid biopsy distinguishes diverse blood based evolving biomarkers comprising circulating tumor cells (CTCs), circulating tumor DNA (ctDNA) or cfDNA, circulating RNA (cfRNA) and exosomes. Cell free DNA are little DNA fragments found circulating in plasma or serum, just as other fluids present in our body. Cell free DNA involves primarily double stranded nuclear DNA and mitochondrial DNA, present both on a surface level and in the lumen of vesicles. The probable origins of the tumor-inferred portion of cfDNA are apoptosis or tumor necrosis, lysis of CTCs or release of DNA from the tumor cells into circulation. The evolution of innovations, refinement and improvement in therapeutics for determination of cfDNA fragment size and its distribution provide significant information related with pathological conditions of the cell, thus emerging as promising indicator for clinical output in medical biotechnology.

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Potential clinical applications of circulating cell-free DNA in ovarian cancer patients

Expert Reviews in Molecular Medicine ◽

10.1017/erm.2018.5 ◽

2018 ◽

Vol 20 ◽

Cited By ~ 4

Author(s):

Ana Barbosa ◽

Ana Peixoto ◽

Pedro Pinto ◽

Manuela Pinheiro ◽

Manuel R. Teixeira

Keyword(s):

Ovarian Cancer ◽

Cancer Patients ◽

Liquid Biopsy ◽

Clinical Applications ◽

Epigenetic Alterations ◽

Cell Free Dna ◽

Non Invasive ◽

Free Dna ◽

Potential Use ◽

Circulating Cell Free Dna

AbstractCirculating cell-free DNA (cfDNA) consists of small fragments of DNA that circulate freely in the bloodstream. In cancer patients, a fraction of cfDNA is derived from tumour cells, therefore containing the same genetic and epigenetic alterations, and is termed circulating cell-free tumour DNA. The potential use of cfDNA, the so-called ‘liquid biopsy’, as a non-invasive cancer biomarker has recently received a lot of attention. The present review will focus on studies concerning the potential clinical applications of cfDNA in ovarian cancer patients.

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Detection of Three Distinct Clonal Populations Using Circulating Cell-Free DNA: A Cautionary Note on the Use of Liquid Biopsy

JCO Precision Oncology ◽

10.1200/po.19.00193 ◽

2019 ◽

pp. 1-6

Author(s):

Gregory M. Riedlinger ◽

Nahed Jalloul ◽

Elizabeth Poplin ◽

Janice M. Mehnert ◽

Roman Groisberg ◽

...

Keyword(s):

Liquid Biopsy ◽

Cautionary Note ◽

Cell Free Dna ◽

Free Dna ◽

Circulating Cell Free Dna

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Liquid Biopsy: Comparison of Mutation Detection Methods for Measurement of RET M918T Circulating Cell-Free DNA in Medullary Thyroid Cancer Patients

Cancer Genetics ◽

10.1016/j.cancergen.2016.04.012 ◽

2016 ◽

Vol 209 (6) ◽

pp. 287 ◽

Cited By ~ 1

Author(s):

Caitlin Evers ◽

Dzifa Y. Duose ◽

Meenakshi Mehrotra ◽

Tao Hai ◽

Michal R. Houston ◽

...

Keyword(s):

Thyroid Cancer ◽

Cancer Patients ◽

Liquid Biopsy ◽

Medullary Thyroid Cancer ◽

Mutation Detection ◽

Detection Methods ◽

Cell Free Dna ◽

Medullary Thyroid ◽

Free Dna ◽

Circulating Cell Free Dna

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JCES 01.26 Circulating Cell-Free DNA of Cerebrospinal Fluid May Function as Liquid Biopsy for Leptomeningeal Metastases of ALK Rearrangement NSCLC

Journal of Thoracic Oncology ◽

10.1016/j.jtho.2017.09.313 ◽

2017 ◽

Vol 12 (11) ◽

pp. S1739

Author(s):

Y. Li ◽

B. Jiang ◽

J.-. Yang ◽

X. Zhang ◽

Z. Zhang ◽

...

Keyword(s):

Cerebrospinal Fluid ◽

Liquid Biopsy ◽

Leptomeningeal Metastases ◽

Alk Rearrangement ◽

Cell Free Dna ◽

Free Dna ◽

Circulating Cell Free Dna

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Mass spectrometric based analysis, characterization and applications of circulating cell free DNA isolated from human body fluids

International Journal of Mass Spectrometry ◽

10.1016/j.ijms.2010.10.003 ◽

2011 ◽

Vol 304 (2-3) ◽

pp. 172-183 ◽

Cited By ~ 26

Author(s):

Vaneet K. Sharma ◽

Paul Vouros ◽

James Glick

Keyword(s):

Human Body ◽

Body Fluids ◽

Mass Spectrometric ◽

Cell Free Dna ◽

Free Dna ◽

Circulating Cell Free Dna

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Sensitivity assessment of workflows detecting rare circulating cell-free DNA targets: A study design proposal

PLoS ONE ◽

10.1371/journal.pone.0253401 ◽

2021 ◽

Vol 16 (7) ◽

pp. e0253401

Author(s):

Thorsten Voss ◽

Andrea Ullius ◽

Maike Schönborn ◽

Uwe Oelmüller

Keyword(s):

Study Design ◽

Liquid Biopsy ◽

Limit Of Detection ◽

Test Procedure ◽

Blood Collection ◽

Assay Sensitivity ◽

Cell Free Dna ◽

Free Dna ◽

Sensitivity Assessment ◽

Circulating Cell Free Dna

The field of liquid biopsy has seen extensive growth in recent decades, making it one of the most promising areas in molecular diagnostics. Circulating cell-free DNA (ccfDNA) especially is used as an analyte in a growing number of diagnostic assays. These assays require specified preanalytical workflows delivering ccfDNA in qualities and quantities that facilitate correct and reliable results. As each step and component used in the preanalytical process has the potential to influence the assay sensitivity and other performance characteristics, it is key to find an unbiased experimental setup to test these factors in diagnostic or research laboratories. We defined one such setup by using blood from healthy subjects and commercially available products for blood collection, spike-in material, ccfDNA isolation, and qPCR assays. As the primary read-out, we calculated the probit model-based LOD95 (limit of detection of the 95th percentile) from the qPCR assay results. In a proof of principle study we tested two different but widely used blood ccfDNA profile stabilization technologies in blood collection tubes, the Cell-Free DNA BCT and the PAXgene Blood ccfDNA Tube. We tested assays for three different EGFR gene mutations and one BRAF gene mutation. The study design revealed differences in performance between the two tested technologies for all four mutations. In conclusion, we successfully established a blueprint for a test procedure capable of verifying and validating a liquid biopsy workflow from blood collection to the analytical result.

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Circulating Cell-Free DNA-Based Liquid Biopsy Markers for the Non-Invasive Prognosis and Monitoring of Metastatic Pancreatic Cancer

Cancers ◽

10.3390/cancers12071754 ◽

2020 ◽

Vol 12 (7) ◽

pp. 1754 ◽

Cited By ~ 1

Author(s):

Marta Toledano-Fonseca ◽

M. Teresa Cano ◽

Elizabeth Inga ◽

Rosa Rodríguez-Alonso ◽

M. Auxiliadora Gómez-España ◽

...

Keyword(s):

Liquid Biopsy ◽

Hepatic Metastases ◽

Progression Free Survival ◽

The Body ◽

Ductal Adenocarcinoma ◽

Cell Free Dna ◽

Ras Mutation ◽

Non Invasive ◽

Free Dna ◽

Circulating Cell Free Dna

Liquid biopsy may assist in the management of cancer patients, which can be particularly applicable in pancreatic ductal adenocarcinoma (PDAC). In this study, we investigated the utility of circulating cell-free DNA (cfDNA)-based markers as prognostic tools in metastatic PDAC. Plasma was obtained from 61 metastatic PDAC patients, and cfDNA levels and fragmentation were determined. BEAMing technique was used for quantitative determination of RAS mutation allele fraction (MAF) in cfDNA. We found that the prognosis was more accurately predicted by RAS mutation detection in plasma than in tissue. RAS mutation status in plasma was a strong independent prognostic factor for both overall survival (OS) and progression-free survival (PFS). Moreover, RAS MAF in cfDNA was also an independent risk factor for poor OS, and was strongly associated with primary tumours in the body/tail of the pancreas and liver metastases. Higher cfDNA levels and fragmentation were also associated with poorer OS and shorter PFS, body/tail tumors, and hepatic metastases, whereas cfDNA fragmentation positively correlated with RAS MAF. Remarkably, the combination of CA19-9 with MAF, cfDNA levels and fragmentation improved the prognostic stratification of patients. Furthermore, dynamics of RAS MAF better correlated with patients’ outcome than standard CA19-9 marker. In conclusion, our study supports the use of cfDNA-based liquid biopsy markers as clinical tools for the non-invasive prognosis and monitoring of metastatic PDAC patients.

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