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

• Published in: PloS one Vol. 9; no. 11; p. e112293
• Main Authors: Malentacchi, Francesca, Pazzagli, Mario, Simi, Lisa, Orlando, Claudio, Wyrich, Ralf, Günther, Kalle, Verderio, Paolo, Pizzamiglio, Sara, Ciniselli, Chiara Maura, Zhang, Hui, Korenková, Vlasta, Rainen, Lynne, Bar, Tzachi, Kubista, Mikael, Gelmini, Stefania
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 10-11-2014; Public Library of Science (PLoS)
• Subjects: Bioinformatics; Biology and life sciences; Biometrics; Blood; Blood & organ donations; Blood Specimen Collection - methods; Collection; Deoxyribonucleic acid; DNA; FosB protein; Gene expression; Gene Expression Profiling; Glyceraldehyde-3-phosphate dehydrogenase; GPI-Linked Proteins - genetics; Humans; Interleukin 1; Interleukin 8; Interleukin-1beta - genetics; Laboratories; Mathematical analysis; Medical statistics; Proto-Oncogene Proteins c-fos - genetics; Quality; Quality assessment; Quality Control; Receptors, Tumor Necrosis Factor, Member 10c; Ribonucleic acid; RNA; RNA - blood; RNA - genetics; Shipping; Shipping industry; Stability analysis; Standardization; Storage; Storage temperature; Tubes; Tumor Necrosis Factor Decoy Receptors - genetics; Italy; Czech Republic; Germany
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0112293

One purpose of the EC funded project, SPIDIA, is to develop evidence-based quality guidelines for the pre-analytical handling of blood samples for RNA molecular testing. To this end, two pan-European External Quality Assessments (EQAs) were implemented. Here we report the results of the second SPIDIA-RNA EQA. This second study included modifications in the protocol related to the blood collection process, the shipping conditions and pre-analytical specimen handling for participants. Participating laboratories received two identical proficiency blood specimens collected in tubes with or without an RNA stabilizer. For pre-defined specimen storage times and temperatures, laboratories were asked to perform RNA extraction from whole blood according to their usual procedure and to return extracted RNA to the SPIDIA facility for further analysis. These RNA samples were evaluated for purity, yield, integrity, stability, presence of interfering substances, and gene expression levels for the validated markers of RNA stability: FOS, IL1B, IL8, GAPDH, FOSB and TNFRSF10c. Analysis of the gene expression results of FOS, IL8, FOSB, and TNFRSF10c, however, indicated that the levels of these transcripts were significantly affected by blood collection tube type and storage temperature. These results demonstrated that only blood collection tubes containing a cellular RNA stabilizer allowed reliable gene expression analysis within 48 h from blood collection for all the genes investigated. The results of these two EQAs have been proposed for use in the development of a Technical Specification by the European Committee for Standardization.