Improvement of molecular monitoring of residual disease in leukemias by bedside RNA stabilization.
Author(s): Müller MC, Merx K, Weisser A, Kreil S, Lahaye T, Hehlmann R, Hochhaus A
Publication: Leukemia, 2002, Vol. 16, Page 2395-9
PubMed ID: 12454744 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
The purpose of this study was to determine if a room temperature delay of 72 h prior to processing of peripheral blood specimens impacts the expression of therapy efficacy genetic markers, and whether a novel RNA stabilization system can attenuate potential effects.
Summary of Findings:
Expression ratios were poorly correlated in unstabilized controls when specimens stored for 2 h and 72 h at room temperature were compared (r=0.65-0.75), indicating a notable effect of storage duration on gene expression; effects were partially attenuated in PAXgene Blood RNA stabilized specimens, generating strong correlations among specimens stored for 2 h and 72 h prior to processing (r=0.95-0.99). Of note, gene expression was significantly higher in specimens extracted using the PAXgene Blood RNA kit compared to RNAeasy Mini Kit processed controls, preventing a direct comparison of stabilization and analyte extraction methods. The sensitivity of the PAXgene Blood RNA stabilization method was comparable to CsCl gradient ultra centrifugation.
Biospecimens
Preservative Types
- None (Fresh)
- PAXgene
Diagnoses:
- Neoplastic - Leukemia
Platform:
Analyte Technology Platform RNA Real-time qRT-PCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Time at room temperature 2 h
72 h
Analyte Extraction and Purification Analyte isolation method PAXgene Blood RNA Kit
RNeasy Mini Kit
Real-time qRT-PCR Specific Targeted nucleic acid BCR-ABL
Total ABL
G6PD
Biospecimen Preservation Type of fixation/preservation PAXgene
None (fresh)
Biospecimen Acquisition Type of collection container/solution PAXgene tube
EDTA Vacutainer