ArticleBMC biotechnology2018
Two approaches for estimating the lower limit of quantitation (LLOQ) of microRNA levels assayed as exploratory biomarkers by RT-qPCR.
Article in BMC biotechnology, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed.
- STALARD: Selective Target Amplification for Low-Abundance RNA Detection.Plant methods · 2025Article
- Universal ddPCR-based assay for the determination of lentivirus infectious titer and lenti-modified cell vector copy number.Molecular therapy. Methods & clinical development · 2023Article
- Harnessing the LdCsm RNA Detection Platform for Efficient microRNA Detection.International journal of molecular sciences · 2023Article
- Confidence interval comparison: Precision of maximum likelihood estimates in LLOQ affected data.PloS one · 2023Article
- Article
- Analyzing siRNA Concentration, Complexation and Stability in Cationic Dendriplexes by Stem-Loop Reverse Transcription-qPCR.Pharmaceutics · 2022Article
- Association between miR-146a and Tumor Necrosis Factor Alpha (TNF-α) in Stable Coronary Artery Disease.Medicina (Kaunas, Lithuania) · 2021Article
- Methodological considerations for measuring biofluid-based microRNA biomarkers.Critical reviews in toxicology · 2021Article
- Cigarette Smoking, miR-27b Downregulation, and Peripheral Artery Disease: Insights into the Mechanisms of Smoking Toxicity.Journal of clinical medicine · 2021Article
- Circulating miRNAs Are Associated with the Systemic Extent of Atherosclerosis: Novel Observations for miR-27b and miR-146.Diagnostics (Basel, Switzerland) · 2021Article
- Deciphering the molecular mechanisms of mother-to-egg immune protection in the mealworm beetle Tenebrio molitor.PLoS pathogens · 2020Article
- Article
- Analytics of Cerebrospinal Fluid MicroRNA Quantitative PCR Studies.Molecular neurobiology · 2019Article
- Validation of bioanalytical method for quantification of Vitamin K2 (MK-4) in human plasma by high-performance liquid chromatography-ultraviolet.Journal of advanced pharmaceutical technology & researchArticle
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15 authors.
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Abstract
backgroundCirculating microRNAs are undergoing exploratory use as safety biomarkers in drug development. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) is one common approach used to quantitate levels of microRNAs in samples that includes the use of a standard curve of calibrators fit to a regression model. Guidelines are needed for setting assay quantitation thresholds that are appropriate for this method and to biomarker pre-validation.
resultsIn this report, we develop two workflows for determining a lower limit of quantitation (LLOQ) for RT-qPCR assays of microRNAs in exploratory studies. One workflow is based on an error threshold calculated by a logistic model of the calibration curve data. The second workflow is based on a threshold set by the sample blank, which is the no template control for RT-qPCR. The two workflows are used to set lower thresholds of reportable microRNA levels for an example dataset in which miR-208a levels in biofluids are quantitated in a cardiac injury model. LLOQ thresholds set by either workflow are effective in filtering out microRNA values with large uncertainty estimates.
conclusionsTwo workflows for LLOQ determinations are presented in this report that provide methods that are easy to implement in investigational studies of microRNA safety biomarkers and offer choices in levels of conservatism in setting lower limits of acceptable values that facilitate interpretation of results.
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