Fauth, Maria ; Hegewald, Anett B. ; Schmitz, Lisa ; Krone, Dorothee J. ; Saul, Meike J. (2024)
Validation of extracellular miRNA quantification in blood samples using RT‐qPCR.
In: FASEB BioAdvances, 2019, 1 (8)
doi: 10.26083/tuprints-00016719
Artikel, Zweitveröffentlichung, Verlagsversion
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Kurzbeschreibung (Abstract)
Extracellular microRNAs (miRs) have been proposed as important blood‐based biomarkers for several diseases. Contrary to proteins and other RNA classes, miRs are stable and easily detectable in body fluids. In this respect, miRs represent a perfect candidate for minimal invasive biomarkers which can hopefully become a complement for invasive histological examinations of tumor tissue. Despite the high number of miR biomarker studies, the specificity and reproducibility of these studies is missing. Therefore, the standardization of pre‐analytical and analytical methods is urgently needed. Here, we validated miR analysis for RNA isolation and miR quantification by quantitative polymerase chain reaction (RT‐qPCR) based on good laboratory practice (GLP). Validation was carried out exemplarily on four miRs, which had already been described as potential biomarkers in previous studies. As basis for RNA analysis using RT‐qPCR, the Minimum Information for Publication of Quantitative Real‐Time PCR Experiments were applied and adapted on the analysis of circulating miRs from human plasma. In our study, we identified and solved several pitfalls from handling to normalization strategy in the analysis of extracellular miRs that lead to inconsistent and non‐repeatable data. Principles of GLP set a framework of experimental design, performance and monitoring to ensure high quality and reliable data. Within this study, we appointed first acceptance criteria for circulating miR quantification during validation which set standards for future miR quantification in blood samples.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2024 |
| Autor(en): | Fauth, Maria ; Hegewald, Anett B. ; Schmitz, Lisa ; Krone, Dorothee J. ; Saul, Meike J. |
| Art des Eintrags: | Zweitveröffentlichung |
| Titel: | Validation of extracellular miRNA quantification in blood samples using RT‐qPCR |
| Sprache: | Englisch |
| Publikationsjahr: | 26 Januar 2024 |
| Ort: | Darmstadt |
| Publikationsdatum der Erstveröffentlichung: | 2019 |
| Ort der Erstveröffentlichung: | Hoboken |
| Verlag: | Wiley |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | FASEB BioAdvances |
| Jahrgang/Volume einer Zeitschrift: | 1 |
| (Heft-)Nummer: | 8 |
| DOI: | 10.26083/tuprints-00016719 |
| URL / URN: | https://tuprints.ulb.tu-darmstadt.de/16719 |
| Zugehörige Links: | |
| Herkunft: | Zweitveröffentlichung DeepGreen |
| Kurzbeschreibung (Abstract): | Extracellular microRNAs (miRs) have been proposed as important blood‐based biomarkers for several diseases. Contrary to proteins and other RNA classes, miRs are stable and easily detectable in body fluids. In this respect, miRs represent a perfect candidate for minimal invasive biomarkers which can hopefully become a complement for invasive histological examinations of tumor tissue. Despite the high number of miR biomarker studies, the specificity and reproducibility of these studies is missing. Therefore, the standardization of pre‐analytical and analytical methods is urgently needed. Here, we validated miR analysis for RNA isolation and miR quantification by quantitative polymerase chain reaction (RT‐qPCR) based on good laboratory practice (GLP). Validation was carried out exemplarily on four miRs, which had already been described as potential biomarkers in previous studies. As basis for RNA analysis using RT‐qPCR, the Minimum Information for Publication of Quantitative Real‐Time PCR Experiments were applied and adapted on the analysis of circulating miRs from human plasma. In our study, we identified and solved several pitfalls from handling to normalization strategy in the analysis of extracellular miRs that lead to inconsistent and non‐repeatable data. Principles of GLP set a framework of experimental design, performance and monitoring to ensure high quality and reliable data. Within this study, we appointed first acceptance criteria for circulating miR quantification during validation which set standards for future miR quantification in blood samples. |
| Freie Schlagworte: | biomarker, GLP, miRNA, plasma, RT‐qPCR, validation |
| Status: | Verlagsversion |
| URN: | urn:nbn:de:tuda-tuprints-167193 |
| Sachgruppe der Dewey Dezimalklassifikatin (DDC): | 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie 600 Technik, Medizin, angewandte Wissenschaften > 610 Medizin, Gesundheit |
| Fachbereich(e)/-gebiet(e): | 10 Fachbereich Biologie 10 Fachbereich Biologie > Extracellular vesicles / miRNA Research |
| Hinterlegungsdatum: | 26 Jan 2024 14:02 |
| Letzte Änderung: | 29 Jan 2024 06:59 |
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| Zugehörige Links: | |
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| Suche nach Titel in: | TUfind oder in Google |
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- Validation of extracellular miRNA quantification in blood samples using RT‐qPCR. (deposited 26 Jan 2024 14:02) [Gegenwärtig angezeigt]
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