Abstract
BackgroundCell-free RNA (cfRNA) naturally occurs in blood and has clinical significance. Accurate quantification of these extracellular RNAs in whole blood is hindered by the simultaneous unintended release of cellular RNA and degradation of cfRNA after blood draw. An appropriate blood collection device is needed to stabilize cfRNA during blood processing, transportation and storage, which will ensure cfRNA test reliability. In this study we compared a novel blood collection device against traditional K3EDTA tubes for its ability to stabilize cfRNA in blood when subjected to conditions that can occur during sample storage and shipping.FindingsShipping blood samples drawn into K3EDTA tubes showed a significant increase in mRNA copy numbers for β-actin, c-fos, and 18S rRNA in plasma. In contrast, shipping blood drawn into Cell-Free RNA BCT™s (BCTs) showed only a slight change in mRNA copy numbers for circulating β-actin, c-fos, and 18S rRNA. Moreover, blood stored in K3EDTA tubes at 6°C, 22°C and 30°C for 3 days showed a significant increase in mRNA copy numbers for c-fos and β-actin, whereas samples stored in BCTs only showed a slight increase.ConclusionOur results show that BCTs minimize increases in background RNA levels caused by temperature fluctuations or agitation that can occur during blood sample storage and shipping. This novel blood collection tube could provide a method for obtaining high quality stabilized cfRNA samples for rare RNA target detection and determining accurate cfRNA concentrations.
Highlights
Cell-free RNA naturally occurs in blood and has clinical significance
Our results show that BCTs minimize increases in background RNA levels caused by temperature fluctuations or agitation that can occur during blood sample storage and shipping
This novel blood collection tube could provide a method for obtaining high quality stabilized Cell-free RNA (cfRNA) samples for rare RNA target detection and determining accurate cfRNA concentrations
Summary
Cell-free RNA (cfRNA) naturally occurs in blood and has clinical significance. Evidence accumulated in recent decades indicates that changes in the levels of circulating nucleic acids are associated with certain disease conditions and could be useful for clinical applications, for cancer patients and pregnant women [2]. Other fetal/placental-specific cfRNAs were reported including human placental lactogen, the β-subunit of human chorionic gonadotropin, and corticotrophinreleasing hormone [9]. These findings have implicated cfRNAs as blood biomarkers for the screening or diagnosis of diseases like cancer and the monitoring of therapy. It provides the opportunity for early, noninvasive prenatal genetic testing
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