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

International Journal of Molecular Sciences
Alison LukTherese M Becker

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 opport...Continue Reading

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Citations

Aug 14, 2018·Expert Review of Molecular Diagnostics·Benjamin J DiPardoJames S Tomlinson
Sep 5, 2019·Expert Review of Molecular Diagnostics·Anson SnowJulie E Lang
Oct 21, 2017·International Journal of Molecular Sciences·Carsten Stephan, Klaus Jung
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Methods Mentioned

BETA
PCR

Software Mentioned

GraphPad
ALS CellCelector
QuantaSoft
GraphPad Prism
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