Evidence map›Paper›PMID 26355751›Full record

ArticlePloS one2015

Cross Platform Standardisation of an Experimental Pipeline for Use in the Identification of Dysregulated Human Circulating MiRNAs.

Helena Kelly, Tim Downing, Nina L Tuite, Terry J Smith, Michael J Kerin, Róisín M Dwyer, Eoin Clancy, Thomas Barry, Kate Reddington

Open access · goldAbstract read
In one paragraph

Article in PloS one, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.

0numbers the graph read from it
0cells of the map it votes in
2citing papers in PubMed
0.6field-weighted citation impact, top 32% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

2 citing papers in PubMed, 7 citations in OpenAlex.

  1. Article
  2. Review
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

9 authors at 2 institutions in 1 country.

Helena KellyNucleic Acid Diagnostics Research Laboratory (NADRL), Microbiology, School of Natural Sciences, National University of Ireland, Galway, Ireland.
Tim DowningSchool of Biotechnology, Dublin City University, Dublin, Ireland.
Nina L TuiteNucleic Acid Diagnostics Research Laboratory (NADRL), Microbiology, School of Natural Sciences, National University of Ireland, Galway, Ireland.
Terry J SmithMolecular Diagnostics Research Group (MDRG), School of Natural Sciences, National University of Ireland, Galway, Ireland; Biomedical Diagnostics Institute (BDI) Programme, National University of Ireland, Galway, Ireland.
Michael J KerinDiscipline of Surgery, School of Medicine, National University of Ireland Galway, Galway, Ireland.
Róisín M DwyerDiscipline of Surgery, School of Medicine, National University of Ireland Galway, Galway, Ireland.
Eoin ClancyMolecular Diagnostics Research Group (MDRG), School of Natural Sciences, National University of Ireland, Galway, Ireland; Biomedical Diagnostics Institute (BDI) Programme, National University of Ireland, Galway, Ireland.
Thomas BarryNucleic Acid Diagnostics Research Laboratory (NADRL), Microbiology, School of Natural Sciences, National University of Ireland, Galway, Ireland.
Kate ReddingtonNucleic Acid Diagnostics Research Laboratory (NADRL), Microbiology, School of Natural Sciences, National University of Ireland, Galway, Ireland.
Ollscoil na Gaillimhe – University of Galway · IEDublin City University · IE

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

introductionMicro RNAs (miRNAs) are a class of highly conserved small non-coding RNAs that play an important part in the post-transcriptional regulation of gene expression. A substantial number of miRNAs have been proposed as biomarkers for diseases. While reverse transcriptase Real-time PCR (RT-qPCR) is considered the gold standard for the evaluation and validation of miRNA biomarkers, small RNA sequencing is now routinely being adopted for the identification of dysregulated miRNAs. However, in many cases where putative miRNA biomarkers are identified using small RNA sequencing, they are not substantiated when RT-qPCR is used for validation. To date, there is a lack of consensus regarding optimal methodologies for miRNA detection, quantification and standardisation when different platform technologies are used. MATERIALS AND

methodsIn this study we present an experimental pipeline that takes into consideration sample collection, processing, enrichment, and the subsequent comparative analysis of circulating small ribonucleic acids using small RNA sequencing and RT-qPCR. RESULTS, DISCUSSION,

conclusionsInitially, a panel of miRNAs dysregulated in circulating blood from breast cancer patients compared to healthy women were identified using small RNA sequencing. MiR-320a was identified as the most dysregulated miRNA between the two female cohorts. Total RNA and enriched small RNA populations (<30 bp) isolated from peripheral blood from the same female cohort samples were then tested for using a miR-320a RT-qPCR assay. When total RNA was analysed with this miR-320a RT-qPCR assay, a 2.3-fold decrease in expression levels was observed between blood samples from healthy controls and breast cancer patients. However, upon enrichment for the small RNA population and subsequent analysis of miR-320a using RT-qPCR, its dysregulation in breast cancer patients was more pronounced with an 8.89-fold decrease in miR-320a expression. We propose that the experimental pipeline outlined could serve as a robust approach for the identification and validation of small RNA biomarkers for disease.

Indexed as

Gene Expression Regulation, NeoplasticBreast NeoplasmsCase-Control StudiesCohort StudiesFemaleGene Expression ProfilingHumansMicroRNAsMolecular Sequence AnnotationReal-Time Polymerase Chain ReactionReference StandardsRNA, NeoplasmSequence Analysis, RNAMicroRNAsRNA, Neoplasm

Identifiers

PMID26355751
PMCPMC4565682
OpenAlexW2341944369

What OpenQuestion holds

Textmetadata
LicenceCC BY
Read underepoch 390

Registered trials

None linked

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.