Analytical and Bioanalytical Chemistry

, Volume 407, Issue 22, pp 6873–6878 | Cite as

Comprehensive evaluation of methods to isolate, quantify, and characterize circulating cell-free DNA from small volumes of plasma

  • Florence Mauger
  • Cécile Dulary
  • Christian Daviaud
  • Jean-François Deleuze
  • Jorg Tost
Note

Abstract

Circulating cell-free DNA (ccfDNA) has great potential for non-invasive diagnostics, and prediction and monitoring of treatment response, but its amount is usually limited. Therefore, the choice of methods to extract and characterize ccfDNA is crucial. In the current study, we performed the most comprehensive comparison of methods for ccfDNA extraction (11 methods), quantification (3 methods), and estimation of the integrity index (2 methods) from small quantities of different kinds of plasma. The QIAamp® Circulating Nucleic Acid Kit and the Norgen Plasma/Serum Circulating DNA Purification Mini Kit showed the best accuracy and reproducibility, but the Norgen kit allowed to extract a higher amount of ccfDNA. This workflow provides a reliable protocol for the multiple applications of ccfDNA in biomedicine.

Graphical Abstract

Workflow for the evaluation of methods to isolate, quantify and characterize circulating cell-free DNA from small volumes of plasma

Keywords

Circulating cell-free DNA Plasma Extraction Quantification Integrity index Cancer Non-invasive diagnostics and personalized treatment 

Notes

Acknowledgments

The authors would like to thank Chao Feng (CNG) for his help with the statistical analysis and Steven McGinn (CNG) for the critical reading of the manuscript. This work was partly supported by the EU FP7 under grant agreement number 241669 (the CAGEKID project, http://www.cng.fr/cagekid).

Supplementary material

216_2015_8846_MOESM1_ESM.pdf (237 kb)
ESM 1(PDF 237 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Florence Mauger
    • 1
  • Cécile Dulary
    • 1
  • Christian Daviaud
    • 1
  • Jean-François Deleuze
    • 1
  • Jorg Tost
    • 1
  1. 1.Laboratory for Epigenetics and Environment, Centre National de GénotypageCEA-Institut de GénomiqueEvryFrance

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