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| Status |
Public on Aug 13, 2021 |
| Title |
Integrating whole blood transcriptomic collection procedures into the current anti-doping testing system, including long-term storage and re-testing of anti-doping samples |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
Recombinant human erythropoietin administration studies involving transcriptomic approaches have demonstrated a gene-expression signature that could aid detection of blood doping. However, current anti-doping testing does not involve blood collection into tubes with RNA preservative. This study investigated if whole blood in long-term storage and whole blood leftover from standard haematological testing in short-term storage could be used for transcriptomic analysis despite lacking RNA preservative. Whole blood samples were collected from thirteen and fourteen healthy males, for long-term and short-term storage experiments. Long-term storage: whole blood collected into Tempus™ tubes and K2EDTA tubes and subjected to long-term (i.e., −80°C) storage and RNA extracted. After storage, K2EDTA tubes were thawed and extracted using GeneJET RNA Purification Kit (Thermo Fisher Scientific, Vilnius, Lithuania) or Tempus™ Spin RNA Isolation Kit (Life Technologies, Carlsbad, CA, USA). RNA quality and purity was sufficient for gene expression analysis. Principle Component Analysis of microarray and RNA-seq gene expression data for long-term storage: When comparing gene expression between blood tubes with and without RNA preservation, 6% (4058 transcripts) were differentially expressed. RNA quantity, purity and integrity was not significantly compromised from long-term storage in blood storage tubes lacking RNA preservative, indicating that transcriptomic analysis could be conducted using anti-doping samples collected or biobanked without RNA preservation.
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| Overall design |
We analyzed whole blood samples with the highest RNA Integrity Number from 8 male healthy subjects using the Affymetrix Human Exon 1.0 ST platform in the long-term study design. Array data was processed by Affymetrix Exon Array Computational Tool. No techinical replicates were performed.
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| Contributor(s) |
Lima G, Kolliari-Turner A, Malinsky FR, Guppy FM, Martin RP, Wang G, Voss SC, Georgakopoulos C, Borrione P, Pigozzi F, Pitsiladis Y |
| Citation(s) |
34765642 |
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| Submission date |
Aug 12, 2021 |
| Last update date |
Nov 18, 2021 |
| Contact name |
Giscard Humberto Oliveira Lima |
| E-mail(s) |
giscard.lima@gmail.com
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| Phone |
+55 11 987353676
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| Organization name |
University of Rome Foro Italico
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| Street address |
Piazza Lauro de Bosis, 15
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| City |
Rome |
| ZIP/Postal code |
00135 |
| Country |
Italy |
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| Platforms (1) |
| GPL17586 |
[HTA-2_0] Affymetrix Human Transcriptome Array 2.0 [transcript (gene) version] |
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| Samples (32)
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| Relations |
| BioProject |
PRJNA754114 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE181970_RAW.tar |
797.4 Mb |
(http)(custom) |
TAR (of CEL, CHP) |
| Processed data included within Sample table |
| Processed data provided as supplementary file |
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