Relevance of pre-analytical blood management on the emerging cardiovascular protein biomarkers TWEAK and HMGB1 and on miRNA serum and plasma profiling

Background: Disease-independent sources of biomarker variability include pre-analytical, analytical and biological variance. The aim of the present study was to evaluate whether the pre-analytical phase has any impact on the emerging heart disease TWEAK and HMGB1 protein markers and miRNA biomarkers...

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Hauptverfasser: Basso, Daniela (VerfasserIn) , Weis, Tanja (VerfasserIn) , Katus, Hugo (VerfasserIn)
Dokumenttyp: Article (Journal)
Sprache:Englisch
Veröffentlicht: 2017
In: Clinical biochemistry
Year: 2016, Jahrgang: 50, Heft: 4, Pages: 186-193
ISSN:1873-2933
DOI:10.1016/j.clinbiochem.2016.11.005
Online-Zugang:Verlag, Volltext: http://dx.doi.org/10.1016/j.clinbiochem.2016.11.005
Verlag, Volltext: http://www.sciencedirect.com/science/article/pii/S0009912016302375
Volltext
Verfasserangaben:Daniela Basso, Andrea Padoan, Thomas Laufer, Vittorio Aneloni, Stefania Moz, Hannah Schroers, Michela Pelloso, Anna Saiz, Medea Krapp, Paola Fogar, Paola Cornoldi, Carlo-Federico Zambon, Elisa Rossi, Marco La Malfa, Alberto Marotti, Thomas Brefort, Tanja M. Weis, Hugo A. Katus, Mario Plebani

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245 1 0 |a Relevance of pre-analytical blood management on the emerging cardiovascular protein biomarkers TWEAK and HMGB1 and on miRNA serum and plasma profiling  |c Daniela Basso, Andrea Padoan, Thomas Laufer, Vittorio Aneloni, Stefania Moz, Hannah Schroers, Michela Pelloso, Anna Saiz, Medea Krapp, Paola Fogar, Paola Cornoldi, Carlo-Federico Zambon, Elisa Rossi, Marco La Malfa, Alberto Marotti, Thomas Brefort, Tanja M. Weis, Hugo A. Katus, Mario Plebani 
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520 |a Background: Disease-independent sources of biomarker variability include pre-analytical, analytical and biological variance. The aim of the present study was to evaluate whether the pre-analytical phase has any impact on the emerging heart disease TWEAK and HMGB1 protein markers and miRNA biomarkers, and whether peptidome profiling allows the identification of pre-analytical quality markers. Methods: An assessment was made of sample type (serum, EDTA-Plasma, Citrate-Plasma, ACD-plasma, Heparin-plasma), temperature of sample storage (room temperature or refrigerated), time of sample storage (0.5, 3, 6 and 9h) and centrifugation (one or two-step). Aliquots of all processed samples were immediately frozen (−80°C) before analysis. Proteins were assayed by ELISAs, miRNA expression profile by microarray and peptidome profiling by MALDI-TOF/MS. Results: Temperature, time and centrifugation had no impact on TWEAK and HMGB1 results, which were significantly influenced by matrix type, TWEAK levels being significantly higher (F=194.7, p<0.0001), and HMGB1 levels significantly lower (F=36.32, p<0.0001) in serum than in any other plasma type. Unsuitable miRNA results were obtained using Heparin-plasma. Serum miRNA expression profiles depended mainly on temperature, while EDTA-plasma miRNA expression profiles were strongly affected by the centrifugation method used. MALDI-TOF/MS allowed the identification of seven features as indices of pre-analytical serum (m/z at 1206, 1350, 1865 and 2021) or EDTA-plasma (m/z 1897, 2740 and 2917) degradation. Conclusions: Serum and EDTA-plasma allow the analysis of both proteins and miRNA emerging biomarkers of heart diseases. Refrigerated storage prevents an altered miRNA expression profile also in cases of a prolonged time-interval between blood drawing and processing. 
534 |c 2016 
650 4 |a Heart diseases 
650 4 |a HMGB1 
650 4 |a MALDI-TOF/MS 
650 4 |a MicroRNA 
650 4 |a Pre-analytics 
650 4 |a Proteomics 
650 4 |a TWEAK 
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