ReviewMolecular oncology2014
Critical appraisal of quantitative PCR results in colorectal cancer research: can we rely on published qPCR results?
Review in Molecular oncology, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 33 papers, 1 of them a synthesis that pooled 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.
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.
Who cites it
33 citing papers in PubMed, 1 synthesis or guideline pooled it.
- Selection of reference genes for quantitative PCR: identifying reference genes for airway epithelial cells exposed toAmerican journal of physiology. Lung cellular and molecular physiology · 2020Pooled it
- Needle in a Haystack: A Droplet Digital Polymerase Chain Reaction Assay to Detect Rare Helminth Parasites Infecting Natural Host Populations.Molecular ecology resources · 2025Article
- When Two-Fold Is Not Enough: Quantifying Uncertainty in Low-Copy qPCR.International journal of molecular sciences · 2025Article
- Exploring transcriptomic databases to identify and experimentally validate tissue-specific consensus reference gene for gene expression normalization in BALB/c mice acutely exposed to 2,3,7,8-Tetrachlorodibenzo-Current research in toxicology · 2025Article
- Development and validation of a droplet digital PCR assay for sensitive detection and quantification ofFrontiers in plant science · 2025Article
- A Strategy for the Selection of RT-qPCR Reference Genes Based on Publicly Available Transcriptomic Datasets.Biomedicines · 2023Article
- Exogenous spike-in mouse RNAs for accurate differential gene expression analysis in barley using RT-qPCR.Biology methods & protocols · 2023Article
- Innovative GenExpA software for selecting suitable reference genes for reliable normalization of gene expression in melanoma.Scientific reports · 2022Article
- Stroma-derived extracellular vesicle mRNA signatures inform histological nature of prostate cancer.Journal of extracellular vesicles · 2021Article
- The carbon source-dependent pattern of antimicrobial activity and gene expression in Pseudomonas donghuensis P482.Scientific reports · 2021Article
- COVID-19 and Diagnostic Testing for SARS-CoV-2 by RT-qPCR-Facts and Fallacies.International journal of molecular sciences · 2021Article
- Validation and identification of reference genes in Chinese hamster ovary cells for Fc-fusion protein production.Experimental biology and medicine (Maywood, N.J.) · 2020Article
- Pathogenic missense mutation pattern of forkhead box genes in neurodevelopmental disorders.Molecular genetics & genomic medicine · 2019Article
- On resolving ambiguities in microbial community analysis of partial nitritation anammox reactors.Scientific reports · 2019Article
- Estrogen receptor profiles across tissues from male and female Rattus norvegicus.Biology of sex differences · 2019Article
- Improving the standardization of mRNA measurement by RT-qPCR.Biomolecular detection and quantification · 2018Article
- dPCR: A Technology Review.Sensors (Basel, Switzerland) · 2018Review
- While it is not deliberate, much of today's biomedical research contains logical and technical flaws, showing a need for corrective action.International journal of medical sciences · 2018Review
- qPCR primer design revisited.Biomolecular detection and quantification · 2017Review
- Droplet Digital PCR versus qPCR for gene expression analysis with low abundant targets: from variable nonsense to publication quality data.Scientific reports · 2017Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The use of real-time quantitative polymerase chain reaction (qPCR) in cancer research has become ubiquitous. The relative simplicity of qPCR experiments, which deliver fast and cost-effective results, means that each year an increasing number of papers utilizing this technique are being published. But how reliable are the published results? Since the validity of gene expression data is greatly dependent on appropriate normalisation to compensate for sample-to-sample and run-to-run variation, we have evaluated the adequacy of normalisation procedures in qPCR-based experiments. Consequently, we assessed all colorectal cancer publications that made use of qPCR from 2006 until August 2013 for the number of reference genes used and whether they had been validated. Using even these minimal evaluation criteria, the validity of only three percent (6/179) of the publications can be adequately assessed. We describe common errors, and conclude that the current state of reporting on qPCR in colorectal cancer research is disquieting. Extrapolated to the study of cancer in general, it is clear that the majority of studies using qPCR cannot be reliably assessed and that at best, the results of these studies may or may not be valid and at worst, pervasive incorrect normalisation is resulting in the wholesale publication of incorrect conclusions. This survey demonstrates that the existence of guidelines, such as MIQE, is necessary but not sufficient to address this problem and suggests that the scientific community should examine its responsibility and be aware of the implications of these findings for current and future research.
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Registered trials
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.