title: Evaluation of a solid matrix for collection and ambient storage of RNA from whole blood.
abstract: BACKGROUND
  Whole blood gene expression-based molecular diagnostic tests are becoming increasingly available. Conventional tube-based methods for obtaining RNA from whole blood can be limited by phlebotomy, volume requirements, and RNA stability during transport and storage. A dried blood spot matrix for collecting high-quality RNA, called RNA Stabilizing Matrix (RSM), was evaluated against PAXgene® blood collection tubes.
  
  
  METHODS
  Whole blood was collected from 25 individuals and subjected to 3 sample storage conditions: 18 hours at either room temperature (baseline arm) or 37°C, and 6 days at room temperature. RNA was extracted and assessed for integrity by Agilent Bioanalyzer, and gene expression was compared by RT-qPCR across 23 mRNAs comprising a clinical test for obstructive coronary artery disease.
  
  
  RESULTS
  RSM produced RNA of relatively high integrity across the various tested conditions (mean RIN ± 95% CI: baseline arm, 6.92 ± 0.24; 37°C arm, 5.98 ± 0.48; 6-day arm, 6.72 ± 0.23). PAXgene samples showed comparable RNA integrity in both baseline and 37°C arms (8.42 ± 0.17; 7.92 ± 0.1 respectively) however significant degradation was observed in the 6-day arm (3.19 ± 1.32). Gene expression scores on RSM were highly correlated between the baseline and 37°C and 6-day study arms (median r = 0.96, 0.95 respectively), as was the correlation to PAXgene tubes (median r = 0.95, p < 0.001).
  
  
  CONCLUSIONS
  RNA obtained from RSM shows little degradation and comparable RT-qPCR performance to PAXgene RNA for the 23 genes analyzed. Further development of this technology may provide a convenient method for collecting, shipping, and storing RNA for gene expression assays.
authors.affiliation: CardioDx, Inc., 2500 Faber Place, Palo Alto, CA 94303, USA.
authors.forename: Heng
authors.initials: H
authors.lastname: Tao
authors.affiliation: CardioDx, Inc., 2500 Faber Place, Palo Alto, CA 94303, USA.
authors.forename: Philip
authors.initials: P
authors.lastname: Beineke
authors.affiliation: GE Global Research, One Research Circle, K1 5D29 Niskayuna NY 12309, USA.
authors.forename: Bing
authors.initials: B
authors.lastname: Li
authors.affiliation: GE Global Research, One Research Circle, K1 5D29 Niskayuna NY 12309, USA.
authors.forename: William
authors.initials: W
authors.lastname: Alberts
authors.affiliation: CardioDx, Inc., 2500 Faber Place, Palo Alto, CA 94303, USA.
authors.forename: Steven
authors.initials: S
authors.lastname: Rosenberg
authors.affiliation: GE Global Research, One Research Circle, K1 5D29 Niskayuna NY 12309, USA.
authors.forename: Erik
authors.initials: E
authors.lastname: Kvam
authors.affiliation: CardioDx, Inc., 2500 Faber Place, Palo Alto, CA 94303, USA.
authors.forename: James A
authors.initials: JA
authors.lastname: Wingrove