| Implementation of a proficiency testing for the assessment of the preanalytical phase of blood RNA. | |
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MedLine Citation:
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PMID: 22285774 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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BACKGROUND: Although important improvements of downstream molecular in vitro diagnostics assays based on RNA from blood were made, the pre-analytical workflow is still poorly defined. METHODS: We performed a multicenter study within the EU-granted SPIDIA project to investigate blood collection and shipping influence on the following RNA quality parameters: yield, purity, integrity, RT-qPCR interference and IL1B, IL8, c-Fos and GAPDH gene expression. Two models were designed: Exp A. Ten laboratories collected blood from an own donor into two different tubes (with or without stabilizer) and extracted RNA at two different times; Exp B. Blood was drawn from a single donor and shipped to ten laboratories in two different tubes (with or without stabilizer) for RNA extraction. RESULTS: In both models and collection tubes, reliable results were obtained for purity, yield, GAPDH expression, and interferences. A substantial variation in RIN (Exp A) and in transcription levels of IL1β, IL8 and c-Fos (Exp B) was observed for blood collected in unstabilized tubes. Overall the variability was higher among data obtained from unstabilized blood samples. CONCLUSIONS: We defined the experimental setup for a larger ring trial throughout Europe. The chosen downstream analyses verified their potential, serving as adequate markers to test quality of blood RNA. |
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Authors:
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Kalle Günther; Francesca Malentacchi; Paolo Verderio; Sara Pizzamiglio; Chiara Maura Ciniselli; Ales Tichopad; Mikael Kubista; Ralf Wyrich; Mario Pazzagli; Stefania Gelmini |
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Publication Detail:
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Type: JOURNAL ARTICLE Date: 2012-1-21 |
Journal Detail:
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Title: Clinica chimica acta; international journal of clinical chemistry Volume: - ISSN: 1873-3492 ISO Abbreviation: - Publication Date: 2012 Jan |
Date Detail:
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Created Date: 2012-1-30 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 1302422 Medline TA: Clin Chim Acta Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
Copyright Information:
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Copyright © 2012. Published by Elsevier B.V. |
Affiliation:
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QIAGEN GmbH, Hilden, Germany; The SPIDIA Consortium, Germany. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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