SPIDIA-RNA: first external quality assessment for the pre-analytical phase of blood samples used for RNA based analyses.

Author(s): Pazzagli M, Malentacchi F, Simi L, Orlando C, Wyrich R, Günther K, Hartmann CC, Verderio P, Pizzamiglio S, Ciniselli CM, Tichopad A, Kubista M, Gelmini S

Publication: Methods, 2013, Vol. 59, Page 20-31

PubMed ID: 23110812 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 delayed RNA extraction from blood specimens on RNA yield, quality and expression. Blood was collected from 7 healthy volunteers into citrate phosphate dextrose adenine bags, pooled, aliquoted into K2 EDTA or PAXgene tubes and stored overnight at 4 degrees C. Blood specimens were then shipped by international courier throughout Europe on gel packs, and stored at 4 degrees C until extraction. The first extraction was performed within 24 h of arrival at the laboratory (0 h)and the second was performed 24 h after the first extraction. RNA was shipped back on dry ice to SPIDIA and stored at -80 degrees C until analysis. Results were not compared between preservative systems, nor were details of extraction methods presented.

   Summary of Findings:

   The distribution of yields was broader and the variability in RINs was higher among specimens extracted 24 h after arrival in the laboratory than in those extracted immediately after arrival, but the median yields, RINs and gene expression profiles were similar between the specimens. Fewer GAPDH copies were quantified by real-time RT-PCR in specimens with a RIN <5 than in specimens with a RIN >5.

   Biospecimens

   • Bodily Fluid - Blood

   Preservative Types

   • Other Preservative
   • PAXgene

   Diagnoses:

   • Normal

   Platform:

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

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	PAXgene Refrigeration
   Real-time qRT-PCR Specific	Targeted nucleic acid	c-fos IL-1 beta IL-8 GAPDH
   Storage	Storage duration	0 h 24 h

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