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Updated: Jul 7, 2025

A Standardized Liquid Biopsy Preanalytical Protocol for Downstream Circulating-Free DNA Applications
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Liquid biopsy for precision diagnostics and therapeutics.

Kuttiappan Anitha1, Bhargavi Posinasetty2, K Naveen Kumari3

  • 1Department of Pharmacology, School of Pharmacy and Technology Management (SPTM), SVKM's Narsee Monjee Institute of Management Studies (NMIMS) Deemed-to-University, Shirpur 425405, India.

Clinica Chimica Acta; International Journal of Clinical Chemistry
|December 27, 2023
PubMed
Summary

Liquid biopsy offers a non-invasive method for cancer diagnosis by analyzing biomarkers like circulating tumor DNA (ctDNA) in bodily fluids. This approach aids in early detection, treatment monitoring, and understanding tumor characteristics.

Keywords:
BiomarkersCell-free nucleic acids (cfNAs)Circulating tumor DNA (ctDNA)Circulating tumor cells (CTCs)ExosomesLiquid biopsyOncology

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Area of Science:

  • Oncology
  • Molecular Diagnostics
  • Biochemistry

Background:

  • Liquid biopsy (LB) is a minimally invasive diagnostic technique analyzing biomolecules in physiological fluids.
  • It offers advantages over traditional tissue biopsies for oncology and other medical fields.
  • Biomolecules analyzed include circulating tumor DNA (ctDNA), circulating tumor cells (CTCs), and cell-free DNA (cfDNA).

Purpose of the Study:

  • To review current liquid biopsy technologies and clinical applications.
  • To emphasize the isolation and analysis of ctDNA and/or cfDNA.
  • To identify tumor markers for cancer detection, prognosis, and treatment monitoring.

Main Methods:

  • Review of various LB methodologies including droplet digital PCR, BEAMing, TAm-Seq, CAPP-Seq, WGBS-Seq, WES, and WGS.
  • Analysis of ctDNA and cfDNA isolation and detection techniques.
  • Exploration of circulating tumor cell (CTC) isolation via biomarker-based cell capture (positive and negative enrichment).

Main Results:

  • Liquid biopsy provides insights into tumor characteristics, treatment response, and early disease detection.
  • Diverse technologies exist for analyzing ctDNA, cfDNA, and CTCs.
  • Challenges in sensitivity, specificity, standardization, and interpretability are addressed.

Conclusions:

  • Liquid biopsy is a promising non-invasive tool in oncology.
  • Current technologies enable effective identification of tumor markers.
  • Further standardization and validation are crucial for widespread clinical adoption.