ArticleRNA (New York, N.Y.)2010
High-performance quantification of mature microRNAs by real-time RT-PCR using deoxyuridine-incorporated oligonucleotides and hemi-nested primers.
Article in RNA (New York, N.Y.), 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 34 papers.
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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.
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Who cites it
34 citing papers in PubMed, 76 citations in OpenAlex.
- A selective enrichment and specific probe terminal mediated strategy for highly sensitive detection of microRNAs.Nature communications · 2026Article
- PRICE: direct and robust detection of microRNAs at single-nucleotide resolution.Nature communications · 2026Article
- MicroRNA expression signature as a biomarker in the diagnosis of nodal T-cell lymphomas.Cancer cell international · 2024Article
- Circulating microRNAs can predict chemotherapy-induced toxicities in patients being treated for primary breast cancer.Breast cancer research and treatment · 2023Article
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- The new world of RNA diagnostics and therapeutics.Journal of experimental & clinical cancer research : CR · 2023Article
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- A Combinational Approach for More Efficient miRNA Biosensing.Current genomics · 2022Review
- Identification of microRNAs responsive to shear loading in rat skin.International wound journal · 2022Article
- Experimental MicroRNA Detection Methods.Methods in molecular biology (Clifton, N.J.) · 2022Review
- RepressingeLife · 2021Article
- MicroRNA 219-5p inhibits alveolarization by reducing platelet derived growth factor receptor-alpha.Respiratory research · 2021Article
- Molecular exploration of paediatric intracranial germinomas from multi-ethnic Singapore.BMC neurology · 2020Article
- Immune-Related Circulating miR-125b-5p and miR-99a-5p Reveal a High Recurrence Risk Group of Pancreatic Cancer Patients after Tumor Resection.Applied sciences (Basel, Switzerland) · 2019Article
- miPrimer: an empirical-based qPCR primer design method for small noncoding microRNA.RNA (New York, N.Y.) · 2018Article
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- Regulation of alveolar septation by microRNA-489.American journal of physiology. Lung cellular and molecular physiology · 2016Article
- Pilot Study of Serum MicroRNA-21 as a Diagnostic and Prognostic Biomarker in Egyptian Breast Cancer Patients.Molecular diagnosis & therapy · 2015Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
MicroRNAs are small noncoding RNAs that serve as important regulators of eukaryotic gene expression and are emerging as novel diagnostic and therapeutic targets for human diseases. Robust and reliable detection of miRNAs is an essential step for understanding the functional significance of these small RNAs in both physiological and pathological processes. Existing methods for miRNA quantification rely on fluorescent probes for optimal specificity. In this study, we developed a high-performance real-time reverse transcriptase-polymerase chain reaction (RT-PCR) assay that allows specific and rapid detection of mature miRNAs using a fast thermocycling profile (10 sec per cycle). This assay exhibited a wide dynamic range (>7 logs) and was capable of detecting miRNAs from as little as 1 pg of the total RNA or as few as 10 cells. The use of modified reverse-transcription oligonucleotides with a secondary structure and hemi-nested reverse PCR primers allowed excellent discrimination of mature miRNAs from their precursors and highly homologous family members using SYBR Green I. Using a novel approach involving uracil-DNA glycosylase treatment, we showed that carryover of the reverse transcription oligonucleotide to the PCR can be successfully eliminated and discrimination between miRNA homologs could be further enhanced. These assays were further extended for multiplexed detection of miRNAs directly from cell lysates without laborious total RNA isolation. With the robust performance of these assays, we identified several miRNAs that were regulated by glial cell-line-derived neurotrophic factor in human glioblastoma cells. In summary, this method could provide a useful tool for rapid, robust, and cost-effective quantification of existing and novel miRNAs.
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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.