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• W3208425818 endingPage "114431" @default.
• W3208425818 startingPage "114431" @default.
• W3208425818 abstract "Blood-borne miRNAs serve as disease diagnostic biomarkers and await clinical validation. Here, we evaluated Cel-miR-39-3p and miRNA16-5p as calibrator for the quantification of 15 miRNAs linked to hepatic impairment. We added defined copy numbers of Cel-miR-39-3p to plasma of healthy controls (N = 5) and patient samples undergoing liver resection (N = 51). The miRNAs were isolated according to SOPs and quantified by RT-qPCR using the 2-(ΔΔ-CT)-method. Although miRNA16-5p and the spike-in control behaved similar in qPCR assays (R2 = 0.8591) the spike-in control suffered from high inter-patient variability (median 7.6-fold) and low recoveries (median 5.6%, 95% CI 1.5-11.8%). Adding Cel-miR-39-3p to blood samples prior to RNA-isolation improved the recoveries (median 105.7%; 95% CI 29.9-219.9%), yet the inter-patient variability remained high (median 7.2-fold). Alike, we observed significant variability in CT-values for miRNA16-5p (range 14.7-fold) thus rendering this internal, blood-borne reference gene unacceptable as comparator. Specifically, 10 out of 15 diagnostic miRNAs failed the criteria R2 ≥ 0.8 even though we added a defined copy number of Cel-miR-39-3p. This suggests interference of the spike-in control with individual miRNAs in the assay. Our study highlights current limitations in the quantification of blood-borne miRNAs that is of particularly importance when used for disease diagnostic and therapeutic interventions." @default.
• W3208425818 created "2021-11-08" @default.
• W3208425818 creator A5071725242 @default.
• W3208425818 creator A5089631740 @default.
• W3208425818 date "2021-12-01" @default.
• W3208425818 modified "2023-09-23" @default.
• W3208425818 title "Reliable miRNA biomarker quantification in clinical practice – are we there yet?" @default.
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• W3208425818 doi "https://doi.org/10.1016/j.ab.2021.114431" @default.
• W3208425818 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/34695390" @default.
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