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

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

PLOS ONE, 2017, vol. 12, issue 5, 1-15

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

Date: 2017
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Persistent link: https://EconPapers.repec.org/RePEc:plo:pone00:0177276

DOI: 10.1371/journal.pone.0177276

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