Universität zu Köln

Alidousty, Christina, Brandes, Danielle, Heydt, Carina, Wagener, Svenja, Wittersheim, Maike, Schaefer, Stephan C., Holz, Barbara, Merkelbach-Bruse, Sabine, Buettner, Reinhard, Fassunke, Jana and Schultheis, Anne M. (2017). Comparison of Blood Collection Tubes from Three Different Manufacturers for the Collection of Cell-Free DNA for Liquid Biopsy Mutation Testing. J. Mol. Diagn., 19 (5). S. 801 - 805. NEW YORK: ELSEVIER SCIENCE INC. ISSN 1943-7811

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Abstract

The improvement in sensitive techniques has allowed the detection of tumor-specific aberrations in circulating tumor (ct) DNA. Amplification-refractory mutation system PCR has been used for the analysis of ctDNA to detect resistance-causing mutations in the epidermal growth factor receptor gene (eg, EGFR T790M) in lung cancer patients. So far, Streck tubes have been widely used as standard blood collection tubes. Here, we compared blood collection tubes manufactured by Streck with tubes from Roche and Qiagen regarding their utility in stabilizing ctDNA in plasma samples. Venous blood from healthy donors was collected in tubes from Streck, Roche, and Qiagen. Samples were spiked with artificially fragmented EGFR T790M-mutated DNA and stored for different periods of time or spiked with different DNA amounts before plasma preparation. Extracted ctDNA was analyzed by amplification-refractory mutation system PCR. Mutant DNA, spiked into blood samples from healthy donors at quantities of 1 or 3 ng, was still reliably detectable in all samples after 7 days. EGFR T790M could be detected when spiking was performed with an amount of artificial ctDNA of 0.5 ng when tubes from Roche and Qiagen were used. Blood collection tubes from Roche and Qiagen are highly suitable for ctDNA stabilization and subsequent liquid biopsy testing. Even low ctDNA concentrations allow the detection of somatic mutations.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Alidousty, Christina UNSPECIFIED UNSPECIFIED UNSPECIFIED Brandes, Danielle UNSPECIFIED UNSPECIFIED UNSPECIFIED Heydt, Carina UNSPECIFIED UNSPECIFIED UNSPECIFIED Wagener, Svenja UNSPECIFIED UNSPECIFIED UNSPECIFIED Wittersheim, Maike UNSPECIFIED UNSPECIFIED UNSPECIFIED Schaefer, Stephan C. UNSPECIFIED UNSPECIFIED UNSPECIFIED Holz, Barbara UNSPECIFIED UNSPECIFIED UNSPECIFIED Merkelbach-Bruse, Sabine UNSPECIFIED UNSPECIFIED UNSPECIFIED Buettner, Reinhard UNSPECIFIED UNSPECIFIED UNSPECIFIED Fassunke, Jana UNSPECIFIED UNSPECIFIED UNSPECIFIED Schultheis, Anne M. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-219980
DOI:	10.1016/j.jmoldx.2017.06.004
Journal or Publication Title:	J. Mol. Diagn.
Volume:	19
Number:	5
Page Range:	S. 801 - 805
Date:	2017
Publisher:	ELSEVIER SCIENCE INC
Place of Publication:	NEW YORK
ISSN:	1943-7811
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language CIRCULATING TUMOR DNA; CANCER-PATIENTS; LUNG-CANCER; PLASMA; EGFR; RESISTANCE Multiple languages Pathology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/21998