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© 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.

Abstract

Excellent pre-analytical stability is an essential precondition for reliable molecular profiling of circulating tumor DNA (ctDNA) in oncological diagnostics. Therefore, in vitro degradation of ctDNA and the additional release of contaminating genomic DNA from lysed blood cells must be prevented. Streck Cell-Free DNA blood collection tubes (cfDNA BCTs) have proposed advantages over standard K2EDTA tubes, but mainly have been tested in healthy individuals. Blood was collected from cancer patients (n = 53) suffering from colorectal (n = 21), pancreatic (n = 11), and non-small-cell lung cancer (n = 21) using cfDNA BCT tubes and K2EDTA tubes that were processed immediately or after 3 days (BCTs) or 6 hours (K2EDTA) at room temperature. The cfDNA isolated from these samples was characterized in terms of yield using LINE-1 qPCR; the level of gDNA contamination; and the mutation status of KRAS, NRAS, and EGFR genes using BEAMing ddPCR. CfDNA yield and gDNA levels were comparable in both tube types and were not affected by prolonged storage of blood samples for at least 3 days in cfDNA BCTs or 6 hours in K2EDTA tubes. In addition, biospecimens collected in K2EDTA tubes and cfDNA BCTs stored for up to 3 days demonstrated highly comparable levels of mutational load across all respective cancer patient cohorts and a wide range of concentrations. Our data support the applicability of clinical oncology specimens collected and stored in cfDNA BCTs for up to 3 days for reliable cfDNA and mutation analyses.

Details

Title
Pre-Analytical Evaluation of Streck Cell-Free DNA Blood Collection Tubes for Liquid Profiling in Oncology
Author
Inga Medina Diaz 1 ; Nocon, Annette 1 ; Held, Stefanie A E 2 ; Kobilay, Makbule 3 ; Skowasch, Dirk 4 ; Bronkhorst, Abel J 5 ; Ungerer, Vida 5 ; Fredebohm, Johannes 1 ; Diehl, Frank 1 ; Holdenrieder, Stefan 6   VIAFID ORCID Logo  ; Holtrup, Frank 1 

 Research and Development, Sysmex Inostics GmbH, 20251 Hamburg, Germany 
 Department of Hematology and Oncology, University Hospital, 53127 Bonn, Germany 
 Institute of Clinical Chemistry and Clinical Pharmacology, University Hospital, 53127 Bonn, Germany 
 Department of Pneumology, University Hospital, 53127 Bonn, Germany 
 German Heart Centre Munich, Institute for Laboratory Medicine, Technical University, 80636 Munich, Germany 
 Institute of Clinical Chemistry and Clinical Pharmacology, University Hospital, 53127 Bonn, Germany; German Heart Centre Munich, Institute for Laboratory Medicine, Technical University, 80636 Munich, Germany 
First page
1288
Publication year
2023
Publication date
2023
Publisher
MDPI AG
e-ISSN
20754418
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
2799567637
Copyright
© 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.