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Detection of Cell-Free DNA in Blood Plasma Samples of Cancer Patients
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Circulating cell-free DNA in cancer.

Pamela Pinzani1, Francesca Salvianti, Claudio Orlando

  • 1Department of Biomedical, Experimental and Clinical Sciences, University of Florence, Viale Pieraccini 6, Florence 50139, Italy, p.pinzani@dfc.unifi.it.

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PubMed
Summary

This study addresses challenges in cell-free DNA (cfDNA) analysis, focusing on sample collection and extraction. It presents a method for precise quantification of total cfDNA and tumor-derived cfDNA in plasma.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Genetics

Background:

  • Preanalytical and analytical phases are critical in cell-free DNA (cfDNA) analysis.
  • Existing methods face challenges in sample collection and extraction, impacting cfDNA quantification.
  • Accurate measurement of cfDNA is essential for various diagnostic applications.

Purpose of the Study:

  • To critically evaluate the preanalytical and analytical phases of cfDNA analysis.
  • To propose a robust method for quantifying total cfDNA in plasma.
  • To detail an approach for measuring tumor-derived cfDNA.

Main Methods:

  • Review of existing literature and methodologies for cfDNA analysis.
  • Development and validation of a novel cfDNA quantification assay.
  • Specific protocol for isolating and measuring tumor-derived cfDNA from plasma samples.

Main Results:

  • Identified key areas of criticism in current sample collection and extraction protocols.
  • Demonstrated accurate quantification of total cfDNA in plasma using the described method.
  • Successfully applied the approach to measure tumor-derived cfDNA.

Conclusions:

  • The proposed method enhances the accuracy of cfDNA quantification.
  • Addressing preanalytical and analytical challenges is crucial for reliable cfDNA analysis.
  • This work contributes to improved diagnostic strategies utilizing cfDNA.