ArticlePloS one2016
Evaluation of Existing Methods for Human Blood mRNA Isolation and Analysis for Large Studies.
Article in PloS one, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 22 citations in OpenAlex.
- Unlocking the power of non-coding RNAs: toward real-time cancer monitoring in precision oncology.Molecular cancer · 2026Review
- A method for measuring mitochondrial DNA copy number in pediatric populations.Frontiers in pediatrics · 2024Article
- Whole blood RNA extraction efficiency contributes to variability in RNA sequencing data sets.PloS one · 2023Article
- Development of a neonatal Göttingen Minipig model for dose precision in perinatal asphyxia: technical opportunities, challenges, and potential further steps.Frontiers in pediatrics · 2023Review
- Comparative transcriptomic analysis reveals translationally relevant processes in mouse models of malaria.eLife · 2022Article
- Commercially Available Blocking Oligonucleotides Effectively Suppress Unwanted Hemolysis-Related miRNAs in a Large Whole-Blood RNA Cohort.The Journal of molecular diagnostics : JMD · 2021Article
- Evaluation of FKBP5 as a cortisol activity biomarker in patients with ACTH-dependent Cushing syndrome.Journal of clinical & translational endocrinology · 2021Article
- Detection of remaining Plasmodium DNA and gametocytes during follow up after curative malaria treatment among returned travellers in Norway.Malaria journal · 2020Article
- Evaluation of RNA purification methods by using different blood stabilization tubes: identification of key features for epidemiological studies.BMC research notes · 2020Article
- Comparable type I interferon score determination from PAXgene and Tempus whole blood RNA collection and isolation systems.BMC research notes · 2019Article
- Gene expression profiling of whole blood: A comparative assessment of RNA-stabilizing collection methods.PloS one · 2019Article
- Clinical utility of circulating non-coding RNAs - an update.Nature reviews. Clinical oncology · 2018Review
- Impact of blood collection and processing on peripheral blood gene expression profiling in type 1 diabetes.BMC genomics · 2017Article
Corrections and comments
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Authors and funding
7 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
aimsPrior to implementing gene expression analyses from blood to a larger cohort study, an evaluation to set up a reliable and reproducible method is mandatory but challenging due to the specific characteristics of the samples as well as their collection methods. In this pilot study we optimized a combination of blood sampling and RNA isolation methods and present reproducible gene expression results from human blood samples.
methodsThe established PAXgeneTM blood collection method (Qiagen) was compared with the more recent TempusTM collection and storing system. RNA from blood samples collected by both systems was extracted on columns with the corresponding Norgen and PAX RNA extraction Kits. RNA quantity and quality was compared photometrically, with Ribogreen and by Real-Time PCR analyses of various reference genes (PPIA, β-ACTIN and TUBULIN) and exemplary of SIGLEC-7.
resultsCombining different sampling methods and extraction kits caused strong variations in gene expression. The use of PAXgeneTM and TempusTM collection systems resulted in RNA of good quality and quantity for the respective RNA isolation system. No large inter-donor variations could be detected for both systems. However, it was not possible to extract sufficient RNA of good quality with the PAXgeneTM RNA extraction system from samples collected by TempusTM collection tubes. Comparing only the Norgen RNA extraction methods, RNA from blood collected either by the TempusTM or PAXgeneTM collection system delivered sufficient amount and quality of RNA, but the TempusTM collection delivered higher RNA concentration compared to the PAXTM collection system. The established Pre-analytix PAXgeneTM RNA extraction system together with the PAXgeneTM blood collection system showed lowest CT-values, i.e. highest RNA concentration of good quality. Expression levels of all tested genes were stable and reproducible.
conclusionsThis study confirms that it is not possible to mix or change sampling or extraction strategies during the same study because of large variations of RNA yield and expression levels.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.