Circulating tumour cell RNA characterisation from colorectal cancer patient blood after inertial microfluidic enrichment

Marnie Winter, Zhen Cai, Katharina Winkler, Kristen Georgiou, Daniel Inglis, Tina Lavranos, Meysam Rezaei, Majid Warkiani, Benjamin Thierry

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Abstract

The detection and molecular analysis of circulating tumour cells (CTCs) potentially provides a significant insight to the characterisation of disease, stage of progression and therapeutic options for cancer patients. Following on from the protocol by Warkiani et al. 2016, which describes a method of enriching CTCs from cancer patient blood with inertial microfluidics, we describe a method to measure the CTC RNA expression in the enriched fraction using droplet digital PCR and compare transcript detection with and without RNA pre-amplification. • Inertial microfluidics combined with droplet digital PCR is advantageous as it allows for CTC enrichment and subsequent RNA analysis from patient blood. This allows for patient tumour analysis with increased sensitivity and precision compared to quantitative Real Time PCR and enables the direct quantification of nucleic acids without the need for tumour biopsy. • This method demonstrates an efficient approach providing important insights into the analysis of colorectal cancer patients’ CTCs using a specific gene subset or biomarkers, an approach that may be tailored to tumour type or expanded to larger panels.

Original languageEnglish
Pages (from-to)1512-1520
Number of pages9
JournalMethodsX
Volume6
DOIs
Publication statusPublished - 17 Jun 2019
Externally publishedYes

Bibliographical note

(CC-BY 4.0) Open Access article licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license (http://creativecommons.org/licenses/by/4.0) .

Keywords

  • Circulating tumour cell
  • Circulating tumour cells isolation and analysis with droplet digital PCR
  • Colorectal cancer
  • Droplet digital PCR

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  • Cite this

    Winter, M., Cai, Z., Winkler, K., Georgiou, K., Inglis, D., Lavranos, T., Rezaei, M., Warkiani, M., & Thierry, B. (2019). Circulating tumour cell RNA characterisation from colorectal cancer patient blood after inertial microfluidic enrichment. MethodsX, 6, 1512-1520. https://doi.org/10.1016/j.mex.2019.06.012