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Updated: May 16, 2026
Serum and Plasma Copy Number Detection Using Real-time PCR
Published on: December 15, 2017
Quantification of breast cancer cells in peripheral blood samples by real-time rt-PCR
Michael Zebisch1, Alexandra C Kölbl, Christian Schindlbeck
1Department of Obstetrics and Gynaecology, Ludwig-Maximilians-University of Munich, Maistrasse 11, D-80337 Munich, Germany.
Background:
Circulating tumour cells (CTCs) are cells that have detached from a primary tumour, circulate in the peripheral blood, and are considered to be the main root of distant metastases. We present a method for the detection of CTCs by real-time PCR on different cytokeratin markers.
Materials And Methods:
Blood samples of a healthy donor were mixed with specific numbers of cells from different breast carcinoma cell line cells. RNA was isolated from the samples and transcribed into cDNA. TaqMan real-time PCR for cytokeratins 8, 18 and 19 was carried out and was correlated to that of 18S.
Results:
Cytokeratin gene expression increased in all samples, when as few as 10 tumour cells were added. In the CAMA-1 cell line, the increase was even greater the more cells were added.
Conclusion:
By this methodology, cells from mammary carcinoma cell lines can be detected in blood samples. Its benefit will be validated in samples from patients with breast cancer.
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