Nanopore sequencing from liquid biopsy: analysis of copy number variations from cell-free DNA of lung cancer patients

Mol Cancer. 2021 Feb 12;20(1):32. doi: 10.1186/s12943-021-01327-5.


In the "precision oncology" era the characterization of tumor genetic features is a pivotal step in cancer patients' management. Liquid biopsy approaches, such as analysis of cell-free DNA from plasma, represent a powerful and noninvasive strategy to obtain information about the genomic status of the tumor. Sequencing-based analyses of cell-free DNA, currently performed with second generation sequencers, are extremely powerful but poorly scalable and not always accessible also due to instrumentation costs. Third generation sequencing platforms, such as Nanopore sequencers, aim at overcoming these obstacles but, unfortunately, are not designed for cell-free DNA analysis.Here we present a customized workflow to exploit low-coverage Nanopore sequencing for the detection of copy number variations from plasma of cancer patients. Whole genome molecular karyotypes of 6 lung cancer patients and 4 healthy subjects were successfully produced with as few as 2 million reads, and common lung-related copy number alterations were readily detected.This is the first successful use of Nanopore sequencing for copy number profiling from plasma DNA. In this context, Nanopore represents a reliable alternative to Illumina sequencing, with the advantages of minute instrumentation costs and extremely short analysis time.The availability of protocols for Nanopore-based cell-free DNA analysis will make this analysis finally accessible, exploiting the full potential of liquid biopsy both for research and clinical purposes.

Keywords: Circulating tumor DNA; Copy number aberrations. Diagnosis; Metastasis; Plasma; Third generation sequencing; cfDNA; ctDNA.

Publication types

  • Letter
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Case-Control Studies
  • Cell-Free Nucleic Acids / genetics*
  • DNA Copy Number Variations*
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Liquid Biopsy
  • Lung Neoplasms / diagnosis*
  • Lung Neoplasms / genetics
  • Nanopore Sequencing
  • Sensitivity and Specificity
  • Sequence Analysis, DNA / methods*
  • Workflow


  • Cell-Free Nucleic Acids

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