Assessment of mRNA and microRNA Stabilization in Peripheral Human Blood for Multicenter Studies and Biobanks

Weber, Daniel G.; Casjens, Swaantje; Rozynek, P.; Lehnert, Martin; Zilch-Schöneweis, Sandra; Bryk, Oleksandr; Taeger, Dirk; Gomolka, Maria; Kreuzer, Michaela; Otten, H.; Pesch, Beate; Johnen, Georg; Brüning, Thomas

doi:10.4137/bmi.s5522

Cited by 70 publications

(65 citation statements)

References 33 publications

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“…If high-quality DNA is needed or the target is RNA, samples should be kept frozen at low temperature (see Akor-Dewu et al, 2014;Head et al, 2014) or preserved in RNAlater (e.g. Weber et al, 2010), FTA® cards (e.g. Mendoza et al, 2012) or PAX gene blood tubes until analysis (e.g.…”

Section: Wbcmentioning

confidence: 99%

Inside the Redbox: Applications of haematology in wildlife monitoring and ecosystem health assessment

Maceda‐Veiga

Figuerola

Silvestre³

et al. 2015

Science of The Total Environment

112

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Blood analyses have great potential in studies of ecology, ecotoxicology and veterinary science in wild vertebrates based on advances in human and domestic animal medicine. The major caveat for field researchers, however, is that the 'rules' for human or domestic animal haematology do not always apply to wildlife. The present overview shows the strengths and limitations of blood analyses in wild vertebrates, and proposes a standardisation of pre-analytical procedures plus some suggestions for a more systematic examination of blood smears to increase the diagnostic value of blood data. By discussing the common problems that field researchers face with blood variables, we also aim to highlight common ground enabling new researchers in the field to accurately collect blood samples and interpret and place their haematological findings into the overall picture of an ecological or eco-toxicological study. Besides showing the practicality and ecological relevance of simple blood variables, this study illustrates the suitability of blood samples for the application of cutting-edge analytical procedures for expanding the current repertoire of diagnostic tools in wildlife monitoring and ecosystem health assessment.

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Section: Wbcmentioning

confidence: 99%

Inside the Redbox: Applications of haematology in wildlife monitoring and ecosystem health assessment

Maceda‐Veiga

Figuerola

Silvestre³

et al. 2015

Science of The Total Environment

112

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“…RNAlaterÔ (Protocol C) is an aqueous, nontoxic tissue storage reagent that can stabilize and protect cellular RNA in situ in unfrozen specimens, and can also be used for the processing and preservation of white blood cells, including short-term storage or transportation at room temperature and long-term preservation at -80°C. 13 The PAXgene Blood RNA tube (Protocol F) is a kind of blood collection tube that could stabilize intracellular RNA immediately upon blood collection. Some researchers choose to freeze the blood specimen directly at -80°C (Protocol E) without any additives such stabilizers, to rule out the possibilities of toxicity during long-term storage and interference with other possible future assays.…”

Section: Discussionmentioning

confidence: 99%

Comparison of Six Different Pretreatment Methods for Blood RNA Extraction

Liu

Wang

et al. 2015

Biopreservation and Biobanking

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Human blood specimens serve as important research materials in the field of translational medicine research. The RNA extracted from blood, for example, represents the gene expression profiles of individuals or groups, and can be indicative of the pathological basis for human diseases. Meanwhile, the RNA quality may have severe impacts on the results of RNA studies. RNA is susceptible to many factors, such as the time of sample collection, transportation conditions, protectants, pretreatments, and extraction methods. In this study, six different pretreatment methods are evaluated for their effects on blood RNA extraction including the RNA yields and quality. Results show that most of these methods meet the basic requirements for RNA studies. While considering the simplicity of the procedure, the cost factor, and how to make full use of the samples, the proper method should be employed by researchers who have specific requirements for their research.

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“…In general, miRNA species are much more stable than mRNA and most studies have not used these methods. Several studies have demonstrated successful isolation and quantitation of specific miRNAs or whole miRNomes from whole blood [42][43][44][45] or tissue [46]. However, few studies have systematically compared RNA stabilization media with a control method.…”

Section: Rna Preservation Reagentsmentioning

confidence: 99%

“…One study directly compared Tempus and PAXgene whole blood stabilization systems and identified small differences in miR30b quantitation [43]. Another detected variation in miRNA quantitation isolated from whole blood preserved in either RNAlater or PAXgene, but noted both systems were appropriate for miRNA analysis [45]. A different study of the PAXgene Tissue stabilization system identified high-concordance in miRNA quantitation between frozen and stabilized specimens (R 2 = 0.95) and reduced concordance between stabilized and formalin-fixed, paraffin embedded (FFPE) tissues (R 2 = 0.81).…”

Section: Rna Preservation Reagentsmentioning

confidence: 99%

Variability in, variability out: best practice recommendations to standardize pre-analytical variables in the detection of circulating and tissue microRNAs

Khan

Lieberman

Lockwood

2017

Clinical Chemistry and Laboratory Medicine (CCLM)

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microRNAs (miRNAs) hold promise as biomarkers for a variety of disease processes and for determining cell differentiation. These short RNA species are robust, survive harsh treatment and storage conditions and may be extracted from blood and tissue. Pre-analytical variables are critical confounders in the analysis of miRNAs: we elucidate these and identify best practices for minimizing sample variation in blood and tissue specimens. Preanalytical variables addressed include patient-intrinsic variation, time and temperature from sample collection to storage or processing, processing methods, contamination by cells and blood components, RNA extraction method, normalization, and storage time/conditions. For circulating miRNAs, hemolysis and blood cell contamination significantly affect profiles; samples should be processed within 2 h of collection; ethylene diamine tetraacetic acid (EDTA) is preferred while heparin should be avoided; samples should be "double spun" or filtered; room temperature or 4 °C storage for up to 24 h is preferred; miRNAs are stable for at least 1 year at -20 °C or -80 °C. For tissuebased analysis, warm ischemic time should be < 1 h; cold ischemic time (4 °C) < 24 h; common fixative used for all specimens; formalin fix up to 72 h prior to processing; enrich for cells of interest; validate candidate biomarkers with in situ visualization. Most importantly, all specimen types should have standard and common workflows with careful documentation of relevant pre-analytical variables.

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Assessment of mRNA and microRNA Stabilization in Peripheral Human Blood for Multicenter Studies and Biobanks

Cited by 70 publications

References 33 publications

Inside the Redbox: Applications of haematology in wildlife monitoring and ecosystem health assessment

Inside the Redbox: Applications of haematology in wildlife monitoring and ecosystem health assessment

Comparison of Six Different Pretreatment Methods for Blood RNA Extraction

Variability in, variability out: best practice recommendations to standardize pre-analytical variables in the detection of circulating and tissue microRNAs

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