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Related Experiment Video

Updated: Jul 12, 2026

Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids
06:53

Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids

Published on: June 8, 2019

Optimized Workflow for Sensitive Detection of Circulating Tumor Cells Using qPCR and Pre-Amplification to Minimize

Eva Obermayr1, Robert Zeillinger2

  • 1Molecular Oncology Group, Department of Obstetrics and Gynecology, Comprehensive Cancer Center, Medical University of Vienna, Vienna, Austria. eva.obermayr@muv.ac.at.

Methods in Molecular Biology (Clifton, N.J.)
|July 9, 2026
PubMed
Summary

Detecting circulating tumor cells (CTCs) is crucial for cancer treatment. This study presents a novel microfluidic enrichment and qPCR method with pre-amplification and contamination controls for sensitive and accurate CTC detection.

Keywords:
Circulating tumor cellsCross-contaminationGene expression analysisLiquid biopsyTargeted pre-amplification

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Microfluidics-based High-throughput Circulating Tumor Cell Sorting and Single-cell Sequencing Technology
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Microfluidics-based High-throughput Circulating Tumor Cell Sorting and Single-cell Sequencing Technology

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Last Updated: Jul 12, 2026

Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids
06:53

Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids

Published on: June 8, 2019

Integration of Wet and Dry Bench Processes Optimizes Targeted Next-generation Sequencing of Low-quality and Low-quantity Tumor Biopsies
13:24

Integration of Wet and Dry Bench Processes Optimizes Targeted Next-generation Sequencing of Low-quality and Low-quantity Tumor Biopsies

Published on: April 11, 2016

Microfluidics-based High-throughput Circulating Tumor Cell Sorting and Single-cell Sequencing Technology
09:45

Microfluidics-based High-throughput Circulating Tumor Cell Sorting and Single-cell Sequencing Technology

Published on: November 14, 2025

Area of Science:

  • Oncology
  • Molecular Biology
  • Biotechnology

Background:

  • Circulating tumor cells (CTCs) provide critical information for cancer treatment and monitoring.
  • Challenges in CTC detection include low abundance, heterogeneity, and contamination from leukocytes.
  • Quantitative PCR (qPCR) is valuable for CTC molecular characterization but requires high sensitivity and specificity.

Purpose of the Study:

  • To develop an enhanced workflow for sensitive and accurate detection of circulating tumor cells (CTCs).
  • To improve CTC detection by combining microfluidic enrichment with a pre-amplified qPCR assay.
  • To minimize false positives and leukocyte contamination in CTC detection.

Main Methods:

  • Development of a workflow integrating microfluidic CTC enrichment with qPCR.
  • Implementation of a pre-amplification protocol to boost signal detection for low-abundance CTCs.
  • Utilized deoxyuridine triphosphate (dUTP) and uracil-N-glycosylase (UNG) in pre-amplification to prevent cross-contamination and false positives.

Main Results:

  • The developed workflow demonstrated enhanced sensitivity for detecting specific gene transcripts from CTCs.
  • The use of dUTP and UNG effectively reduced contamination from leukocytes, ensuring assay specificity.
  • The method provides a robust approach for accurate CTC analysis.

Conclusions:

  • This integrated microfluidic enrichment and pre-amplified qPCR workflow offers a sensitive and accurate method for CTC detection.
  • The contamination control strategy enhances the reliability of CTC molecular profiling.
  • This robust approach holds potential for routine clinical application in cancer management.