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Updated: Jun 13, 2026
Serum and Plasma Copy Number Detection Using Real-time PCR
Published on: December 15, 2017
Clinical evaluation of developed PCR-based method with hydrolysis probes for TOP2A copy number evaluation in breast
Anna Zaczek1, Aleksandra Markiewicz, Janusz Jaśkiewicz
1Department of Medical Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdańsk and Medical University of Gdańsk, Gdańsk, Poland. azaczek@gumed.edu.pl
Objectives:
The objective of this study was to develop a new real time PCR-based method for quantitative detection of topoisomerase II alpha (TOP2A) aberrations and to evaluate its clinical utility in breast cancer.
Design And Methods:
The method applied dually labelled hydrolysis probes and Pfaffl quantification method. The study group consisted of 83 consecutive breast cancer patients.
Results:
In the examined tumour samples median TOP2A gene dosage was 1.08 (range 0.34-7.55). TOP2A amplifications were found in 12 tumours (14.5%), no deletion was detected. Statistically significant positive correlation of TOP2A gene dosage with nodal status, tumour grade, and HER2 protein status was found. TOP2A status also correlated with disease free survival.
Conclusions:
The newly developed real time PCR assay showed to be fast and easy to perform. Determined by the method TOP2A gene dosage was shown to be a potent prognostic factor in breast cancer.
