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Blood collection tube and RNA purification method recommendations for extracellular RNA transcriptome profiling

The exRNAQC Consortium, Jasper Anckaert (UGent) , Francisco Avila Cobos (UGent) , Anneleen Decock (UGent) , Philippe Decruyenaere (UGent) , Jill Deleu (UGent) , Katleen De Preter (UGent) , Olivier De Wever (UGent) , Jilke De Wilde (UGent) , Bert Dhondt, et al.
(2025) NATURE COMMUNICATIONS. 16(1).
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Abstract
Blood-based extracellular RNA (cell-free RNA; exRNA) biomarkers require validated sample collection, processing, and quantification procedures. No study to date has systematically tested pre-analytical variables affecting transcriptome-wide exRNA analysis. By evaluating their impact on deep transcriptome profiling of microRNAs and mRNAs in blood plasma or serum, we compared ten blood collection tubes, three blood processing time intervals, and eight RNA purification methods. In addition, we assessed interactions among a selected pre-analytical variable set, resulting in 456 extracellular transcriptomes. Blood preservation tubes failed to stabilize exRNA and RNA purification methods differed significantly in performance, causing variations in concentration, detected gene numbers, replicability and observed transcriptome complexity. Critical interactions between tubes, purification methods and time intervals were identified. We provide 11 analytical performance metrics for exRNA quantification methods and put forward recommendations for both users and manufacturers of RNA purification methods and blood collection tubes, collectively, essential groundwork for exRNA-based precision medicine applications.
Keywords
CELL-FREE RNA, CIRCULATING MICRORNAS, QUALITY-CONTROL, BIOMARKERS, KNOWLEDGE, MIRBASE, SPEED

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MLA
exRNAQC Consortium, The, et al. “Blood Collection Tube and RNA Purification Method Recommendations for Extracellular RNA Transcriptome Profiling.” NATURE COMMUNICATIONS, vol. 16, no. 1, 2025, doi:10.1038/s41467-025-58607-7.
APA
exRNAQC Consortium, T., Anckaert, J., Avila Cobos, F., Decock, A., Decruyenaere, P., Deleu, J., … Yigit, N. (2025). Blood collection tube and RNA purification method recommendations for extracellular RNA transcriptome profiling. NATURE COMMUNICATIONS, 16(1). https://doi.org/10.1038/s41467-025-58607-7
Chicago author-date
Consortium, The exRNAQC, Jasper Anckaert, Francisco Avila Cobos, Anneleen Decock, Philippe Decruyenaere, Jill Deleu, Katleen De Preter, et al. 2025. “Blood Collection Tube and RNA Purification Method Recommendations for Extracellular RNA Transcriptome Profiling.” NATURE COMMUNICATIONS 16 (1). https://doi.org/10.1038/s41467-025-58607-7.
Chicago author-date (all authors)
exRNAQC Consortium, The, Jasper Anckaert, Francisco Avila Cobos, Anneleen Decock, Philippe Decruyenaere, Jill Deleu, Katleen De Preter, Olivier De Wever, Jilke De Wilde, Bert Dhondt, Thibaut D’huyvetter, Celine Everaert, Carolina Fierro, Hetty Helsmoortel, An Hendrix, Eva Hulstaert, Jan Koster, Scott Kuersten, Tim R. Mercer, Pieter Mestdagh, Annelien Morlion, Nele Nijs, Justine Nuytens, Annouck Philippron, Thomas Piofczyk, Franco Alexander Poma Soto, Kathleen Schoofs, Gary P. Schroth, Olivier Thas, Eveline Vanden Eynde, Jo Vandesompele, Tom Van Maerken, Ruben Van Paemel, Kimberly Verniers, Jasper Verwilt, and Nurten Yigit. 2025. “Blood Collection Tube and RNA Purification Method Recommendations for Extracellular RNA Transcriptome Profiling.” NATURE COMMUNICATIONS 16 (1). doi:10.1038/s41467-025-58607-7.
Vancouver
1.
exRNAQC Consortium T, Anckaert J, Avila Cobos F, Decock A, Decruyenaere P, Deleu J, et al. Blood collection tube and RNA purification method recommendations for extracellular RNA transcriptome profiling. NATURE COMMUNICATIONS. 2025;16(1).
IEEE
[1]
T. exRNAQC Consortium et al., “Blood collection tube and RNA purification method recommendations for extracellular RNA transcriptome profiling,” NATURE COMMUNICATIONS, vol. 16, no. 1, 2025.
@article{01JWKN87EQ5FBKPQXCNV2Q03K0,
  abstract     = {{Blood-based extracellular RNA (cell-free RNA; exRNA) biomarkers require validated sample collection, processing, and quantification procedures. No study to date has systematically tested pre-analytical variables affecting transcriptome-wide exRNA analysis. By evaluating their impact on deep transcriptome profiling of microRNAs and mRNAs in blood plasma or serum, we compared ten blood collection tubes, three blood processing time intervals, and eight RNA purification methods. In addition, we assessed interactions among a selected pre-analytical variable set, resulting in 456 extracellular transcriptomes. Blood preservation tubes failed to stabilize exRNA and RNA purification methods differed significantly in performance, causing variations in concentration, detected gene numbers, replicability and observed transcriptome complexity. Critical interactions between tubes, purification methods and time intervals were identified. We provide 11 analytical performance metrics for exRNA quantification methods and put forward recommendations for both users and manufacturers of RNA purification methods and blood collection tubes, collectively, essential groundwork for exRNA-based precision medicine applications.}},
  articleno    = {{4513}},
  author       = {{exRNAQC Consortium, The and Anckaert, Jasper and Avila Cobos, Francisco and Decock, Anneleen and Decruyenaere, Philippe and Deleu, Jill and De Preter, Katleen and De Wever, Olivier and De Wilde, Jilke and Dhondt, Bert and D'huyvetter, Thibaut and Everaert, Celine and Fierro, Carolina and Helsmoortel, Hetty and Hendrix, An and Hulstaert, Eva and Koster, Jan and Kuersten, Scott and Mercer, Tim R. and Mestdagh, Pieter and Morlion, Annelien and Nijs, Nele and Nuytens, Justine and Philippron, Annouck and Piofczyk, Thomas and Poma Soto, Franco Alexander and Schoofs, Kathleen and Schroth, Gary P. and Thas, Olivier and Vanden Eynde, Eveline and Vandesompele, Jo and Van Maerken, Tom and Van Paemel, Ruben and Verniers, Kimberly and Verwilt, Jasper and Yigit, Nurten}},
  issn         = {{2041-1723}},
  journal      = {{NATURE COMMUNICATIONS}},
  keywords     = {{CELL-FREE RNA,CIRCULATING MICRORNAS,QUALITY-CONTROL,BIOMARKERS,KNOWLEDGE,MIRBASE,SPEED}},
  language     = {{eng}},
  number       = {{1}},
  pages        = {{17}},
  title        = {{Blood collection tube and RNA purification method recommendations for extracellular RNA transcriptome profiling}},
  url          = {{http://doi.org/10.1038/s41467-025-58607-7}},
  volume       = {{16}},
  year         = {{2025}},
}
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