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Succinct workflows for circulating tumor cells after enrichment: From systematic counting to mutational profiling

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  • Victor Chun-Lam Wong
  • Josephine Mun-Yee Ko
  • Chi-Tat Lam
  • Maria Li Lung

Abstract

Purpose: This study aims to establish a highly adaptable workflow downstream of microfluidic enrichment for facilitating systematic CTC enumeration and genetic discovery. Methods: To facilitate CTC enumeration, we established a CK/EPCAM-combined immunostaining strategy and an automated CTC analytical pipeline using an open-source image analyzer. By virtue of this workflow, we conducted a pilot study of 56 cancer patients and 21 healthy individuals using a high-throughput spiral microfluidic chip system. To facilitate genetic discovery of somatic mutations in CTCs, we integrated the CTC enumeration into next-generation sequencing and established a straightforward amplicon library comprising diversifier random sequences to sequence CTC samples. Results: The CTC staining and enumeration workflow achieved 80.4% sensitivity and 85.7% specificity (AUC = 0.87, p = 0.004, power = 0.985), as evaluated by ROC analysis. Univariate and multivariate analysis verified that the CTC (CK/EpCAM+CD45−), but not other cell populations, is a significant and independent biomarker for cancer patients (p

Suggested Citation

  • Victor Chun-Lam Wong & Josephine Mun-Yee Ko & Chi-Tat Lam & Maria Li Lung, 2017. "Succinct workflows for circulating tumor cells after enrichment: From systematic counting to mutational profiling," PLOS ONE, Public Library of Science, vol. 12(5), pages 1-15, May.
    • Handle: RePEc:plo:pone00:0177276
    DOI: 10.1371/journal.pone.0177276
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