Detection and Characterization of Circulating Tumor Cells by Quantitative Real-Time PCR

  • Francesca Salvianti
  • Filomena Costanza
  • Gemma Sonnati
  • Pamela PinzaniEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2065)


We propose two different approaches involving the use of quantitative real-time PCR for the detection or analysis of circulating tumor cells. In one case cells are indirectly identified through the expression of a marker mRNA, while in the other one cells are enriched by size prior to be submitted to mutational analysis for a specific target. Both methods have been successfully applied to the study of circulating melanoma cells.

Key words

CTCs Melanoma RT-qPCR Tyrosinase qPCR Somatic mutation BRAFV600E 


  1. 1.
    Chen L, Bode AM, Dong Z (2017) Circulating tumor cells: moving biological insights into detection. Theranostics 7:2606–2619. Scholar
  2. 2.
    Alix-Panabieres C, Pantel K (2014) Challenges in circulating tumour cell research. Nat Rev Cancer 14:623–631. Scholar
  3. 3.
    Qian W, Zhang Y, Chen W (2015) Capturing cancer: emerging microfluidic technologies for the capture and characterization of circulating tumor cells. Small 11:3850–3872. Scholar
  4. 4.
    Huang L, Bian S, Cheng Y et al (2017) Microfluidics cell sample preparation for analysis: advances in efficient cell enrichment and precise single cell capture. Biomicrofluidics 11:011501. Scholar
  5. 5.
    Rodic S, Mihalcioiu C, Saleh RR (2014) Detection methods of circulating tumor cells in cutaneous melanoma: a systematic review. Crit Rev Oncol Hematol 91:74–92. Scholar
  6. 6.
    Andergassen U, Kölbl AC, Mahner S et al (2016) Real-time RT-PCR systems for CTC detection from blood samples of breast cancer and gynaecological tumour patients (review). Oncol Rep 35:1905–1915. Scholar
  7. 7.
    Yang C, Zou K, Zheng L et al (2017) Prognostic and clinicopathological significance of circulating tumor cells detected by RT-PCR in non-metastatic colorectal cancer: a meta-analysis and systematic review. BMC Cancer 17:725. Scholar
  8. 8.
    Pinzani P, Mazzini C, Salvianti F et al (2010) Tyrosinase mRNA levels in the blood of uveal melanoma patients: correlation with the number of circulating tumor cells and tumor progression. Melanoma Res 20:303–310. Scholar
  9. 9.
    Salvianti F, Orlando C, Massi D et al (2016) Tumor-related methylated cell-free DNA and circulating tumor cells in melanoma. Front Mol Biosci 2:76. Scholar
  10. 10.
    Malentacchi F, Pazzagli M, Simi L et al (2014) SPIDIA-RNA: second external quality assessment for the pre-analytical phase of blood samples used for RNA based analyses. PLoS One 9:e112293. Scholar
  11. 11.
    Desitter I, Guerrouahen BS, Benali-Furet N et al (2011) A new device for rapid isolation by size and characterization of rare circulating tumor cells. Anticancer Res 31:427–441PubMedGoogle Scholar
  12. 12.
    Rainen L, Oelmueller U, Jurgensen S et al (2002) Stabilization of mRNA expression in whole blood samples. Clin Chem 48:1883–1890PubMedGoogle Scholar
  13. 13.
    Pinzani P, Salvianti F, Cascella R et al (2010) Allele specific Taqman-based real-time PCR assay to quantify circulating BRAFV600E mutated DNA in plasma of melanoma patients. Clin Chim Acta 411:1319–1324. Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Francesca Salvianti
    • 1
  • Filomena Costanza
    • 1
  • Gemma Sonnati
    • 1
  • Pamela Pinzani
    • 1
    Email author
  1. 1.Department of Experimental and Clinical Biomedical Sciences “Mario Serio”University of FlorenceFlorenceItaly

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