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Int. J. Mol. Sci. 2017, 18(5), 1047; doi:10.3390/ijms18051047

CTC-mRNA (AR-V7) Analysis from Blood Samples—Impact of Blood Collection Tube and Storage Time

1
Centre for Circulating Tumour Cell Diagnostics and Research, Ingham Institute for Applied Medical Research, 1 Campbell St., Liverpool, NSW 2170, Australia
2
Department of Medical Oncology, Liverpool Hospital, Elizabeth St & Goulburn St, Liverpool, NSW 2170, Australia
3
Western Sydney University Clinical School, Elizabeth St, Liverpool, NSW 2170, Australia
4
South Western Clinical School, University of New South Wales, Goulburn St., Liverpool, NSW 2170, Australia
5
Tokai Pharmaceuticals, Inc., 255 State Street, 6th Floor, Boston, MA 0210, USA
Current address: Siamab Therapeutics, 90 Bridge Street, Suite 100, Newton, MA 02458, USA.
*
Author to whom correspondence should be addressed.
Academic Editor: Carsten Stephan
Received: 31 March 2017 / Revised: 2 May 2017 / Accepted: 8 May 2017 / Published: 12 May 2017
(This article belongs to the Special Issue Diagnostic, Prognostic and Predictive Biomarkers in Prostate Cancer)
View Full-Text   |   Download PDF [2542 KB, uploaded 16 May 2017]   |  

Abstract

Circulating tumour cells (CTCs) are an emerging resource for monitoring cancer biomarkers. New technologies for CTC isolation and biomarker detection are increasingly sensitive, however, the ideal blood storage conditions to preserve CTC-specific mRNA biomarkers remains undetermined. Here we tested the preservation of tumour cells and CTC-mRNA over time in common anticoagulant ethylene-diamine-tetra-acetic acid (EDTA) and acid citrate dextrose solution B (Citrate) blood tubes compared to preservative-containing blood tubes. Blood samples spiked with prostate cancer cells were processed after 0, 24, 30, and 48 h storage at room temperature. The tumour cell isolation efficiency and the mRNA levels of the prostate cancer biomarkers androgen receptor variant 7 (AR-V7) and total AR, as well as epithelial cell adhesion molecule (EpCAM) were measured. Spiked cells were recovered across all storage tube types and times. Surprisingly, tumour mRNA biomarkers were readily detectable after 48 h storage in EDTA and Citrate tubes, but not in preservative-containing tubes. Notably, AR-V7 expression was detected in prostate cancer patient blood samples after 48 h storage in EDTA tubes at room temperature. This important finding presents opportunities for measuring AR-V7 expression from clinical trial patient samples processed within 48 h—a much more feasible timeframe compared to previous recommendations. View Full-Text
Keywords: circulating tumour cell; biomarker; androgen receptor; AR-V7; droplet digital polymerase chain reaction (ddPCR); blood storage tube circulating tumour cell; biomarker; androgen receptor; AR-V7; droplet digital polymerase chain reaction (ddPCR); blood storage tube
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Luk, A.W.S.; Ma, Y.; Ding, P.N.; Young, F.P.; Chua, W.; Balakrishnar, B.; Dransfield, D.T.; Souza, P.; Becker, T.M. CTC-mRNA (AR-V7) Analysis from Blood Samples—Impact of Blood Collection Tube and Storage Time. Int. J. Mol. Sci. 2017, 18, 1047.

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