Automated genotyping of circulating tumor cells

Expert Rev Mol Diagn. 2010 Sep;10(6):723-9. doi: 10.1586/erm.10.66.

Abstract

Cancer remains a prominent health concern in modern societies. Continuous innovations and introduction of new technologies are essential to level or reduce current healthcare spending. A diagnostic platform to detect circulating tumor cells (CTCs) in peripheral blood may be most promising in this respect. CTCs have been proposed as a minimally invasive, prognostic and predictive marker to reflect the biological characteristics of tumors and are implemented in an increasing number of clinical studies. Still, their detection remains a challenge as they may occur at concentrations below one single cell per ml of blood. To facilitate their detection, here we describe microfluidic modules to isolate and genotype CTCs directly from clinical blood samples. In a first cell isolation and detection module, the CTCs are immunomagnetically enriched, separated and counted. In a second module and after cell lysis, the mRNA is reversely transcripted to cDNA, followed by a multiplex ligation probe amplification of 20 specific genetic markers and two control fragments. Following the multiplex ligation probe amplification reaction, the amplified fragments are electrochemically detected in a third and final module. Besides the design of the modules, their functionality is described using control samples. Further testing using clinical samples and integration of all modules in a single, fully automated smart miniaturized system will enable minimal invasive testing for frequent detection and characterization of CTCs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biosensing Techniques
  • Cell Line, Tumor
  • Electrochemical Techniques
  • Genotype*
  • Humans
  • Immunomagnetic Separation / instrumentation
  • Immunomagnetic Separation / methods
  • Neoplasms / blood*
  • Neoplasms / diagnosis
  • Neoplasms / genetics*
  • Neoplastic Cells, Circulating*
  • Nucleic Acid Amplification Techniques