Incubation of whole blood at room temperature does not alter the plasma concentrations of microRNA-16 and -223.
Author(s): Benson EA, Skaar TC
Publication: Drug Metab Dispos, 2013, Vol. 41, Page 1778-81
PubMed ID: 23886700 PubMed Review Paper? No
Purpose of Paper
The purpose of this paper was to investigate the effects of delayed centrifugation of blood on levels of microRNA (miRNA, miR)-16 and -223 in plasma. Inter- and intra-individual variability in miR-16 and miR-223 levels were also investigated.
Conclusion of Paper
There were no consistent or significant changes in levels of miR-16 or miR-223 in plasma after precentrifugation storage of blood for up to 12 h at room temperature. While interindividual variability in cycle threshold values was observed, the coefficient of variance (CV) was similar between patients and intra-individual variability was low.
Studies
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Study Purpose
The purpose of this study was to investigate the effects of delayed centrifugation of blood on levels of miR-16 and miR-223 in plasma. Inter- and intra-individual variability in miR-16 and miR-223 levels were also investigated. Blood was collected twice from four healthy donors into EDTA vacutainer tubes using a 21-gauge needle at an interval of 7-49 days. Blood was inverted eight times and then stored at room temperature for 30 min, 1 h, 2 h, 4 h, 8 h, or 12 h before centrifugation. Plasma was isolated by centrifugation at 1500 x g for 15 min at 4°C, transferred to a new tube, and then stored frozen at -20°C. Plasma was thawed and then recentrifuged at 1500 x g for 10 min at 4°C before isolation of miRNA using the miRNeasy plasma protocol and storage at -80°C. miRNA was quantified by TaqMan real-time PCR Small RNA Assays.
Summary of Findings:
There were no consistent or significant changes in levels of miR-16 or miR-223 in plasma after precentrifugation storage of blood at room temperature. Although the average cycle threshold (CT) values over all timepoints differed among the four subjects by 3-4 CTs, the coefficient of variance (CV) was similar across the subjects for both miR-16 (1.48-4.09%) and miR-224 (3.67 to 6.75%). Levels of miR-16 and miR-223 in immediately processed specimens differed by 0.42 to 7.9% and 1.7 to 8.3%; respectively, between the two blood draws indicating limited intra-individual variability.
Biospecimens
Preservative Types
- Frozen
Diagnoses:
- Normal
Platform:
Analyte Technology Platform RNA Real-time qRT-PCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Aliquots and Components Centrifugation Centrifugation delays investigated
Real-time qRT-PCR Specific Targeted nucleic acid miR-16
miR-223
Storage Storage duration 30 min
1 h
2 h
4 h
8 h
12 h