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?ConclusionRNA 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.

SUBMITTER: Tao H 

PROVIDER: S-EPMC4030268 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature