Assessment of mRNA and microRNA Stabilization in Peripheral Human Blood for Multicenter Studies and Biobanks.

Author(s): Weber DG, Casjens S, Rozynek P, Lehnert M, Zilch-Schöneweis S, Bryk O, Taeger D, Gomolka M, Kreuzer M, Otten H, Pesch B, Johnen G, Brüning T

Publication: Biomark Insights, 2010, Vol. 5, Page 95-102

PubMed ID: 20981139 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of stabilization of RNA in blood with PAXgene or RNAlater during shipping and frozen storage. Peripheral blood was collected from former uranium miners in an EDTA vacutainer, aliquoted, and mixed with RNAlater, or blood was collected directly into PAXgene tubes. RNA was extracted from PAXgene specimens using the PAXgene kit, but RNA was extracted from RNAlater-preserved specimens using the RiboPure blood kit.

   Summary of Findings:

   The RNA integrity and yield were higher from the RNAlater-preserved specimens than from the PAXgene-preserved specimens (p<0.0001), and RNA yield was higher from specimens shipped on dry ice than those shipped at ambient temperatures (p<0.0001). The highest RNA yield and integrity were observed in RNAlater specimens shipped on dry ice and then extracted immediately or stored frozen for 6 months. The lowest RNA yield and integrity were observed in PAXgene specimens shipped at ambient temperatures. Frozen and stored PAXgene-preserved specimens had higher RNA yields than frozen specimens not treated with either PAXgene or RNAlater (2.2 ug/mL versus 1.6 ug/mL; p=0.0002). A slight reduction in RNA yield and integrity were observed in PAXgene specimens shipped at ambient temperature for 4 days compared to 1-2 days, but a similar decrease was not observed in RNAlater-preserved specimens, and there were not enough specimens to determine statistical significance. Relative to GAPDH levels, levels of ATM and miRNA26b transcripts were generally not affected by preservation system or shipping conditions, but significantly more ATM was detected in RNAlater specimens that were shipped on dry ice and stored frozen for 6 months than those shipped frozen and not stored (p<0.0001), and more miRNA26b was detected in PAXgene specimens shipped at ambient temperatures rather than on dry ice (p<0.0001) and in RNAlater, rather than PAXgene-preserved specimens, shipped on dry ice and stored frozen (p<0.0001). In contrast, relative miRNA26a levels were higher in unstored RNAlater-preserved specimens than in PAXgene-preserved specimens (both shipping conditions, p<0.0001) and after frozen storage compared to specimens processed immediately (both preservation systems, p<0.0001).

   Biospecimens

   • Bodily Fluid - Blood

   Preservative Types

   • RNAlater
   • PAXgene

   Diagnoses:

   • Not specified

   Platform:

   Analyte	Technology Platform
   RNA	Automated electrophoresis/Bioanalyzer
   RNA	Real-time qRT-PCR
   RNA	Spectrophotometry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Between site transportation method	Courier Mailed
   Storage	Specimen transport duration/condition	At ambient temperature On dry ice Overnight 1-2 days 4 days
   Storage	Storage duration	0 days 6 months
   Analyte Extraction and Purification	Analyte isolation method	RiboPure blood kit PAXgene blood kit
   Real-time qRT-PCR Specific	Targeted nucleic acid	ATM GAPDH miRNA26a miRNA26b
   Biospecimen Preservation	Type of fixation/preservation	PAXgene RNAlater

   View more