Web of Science: 10 citas, Scopus: 11 citas, Google Scholar: citas,
Practical data handling pipeline improves performance of qPCR-based circulating miRNA measurements
De Ronde, Maurice W.J. (University of Amsterdam)
Ruijter,Jan M. (University of Amsterdam)
Lanfear, David (Henry Ford Hospital. Heart and Vascular Insitute)
Bayes Genis, Antoni (Institut Germans Trias i Pujol. Hospital Universitari Germans Trias i Pujol)
Kok, Maayke G.M. (University of Amsterdam)
Creemers, Esther E. (University of Amsterdam)
Pinto, Yigal M. (University of Amsterdam)
Pinto-Sietsma, Sara-Joan (University of Amsterdam)
Universitat Autònoma de Barcelona

Fecha: 2017
Resumen: Since numerous miRNAs have been shown to be present in circulation, these so-called circulating miRNAs have emerged as potential biomarkers for disease. However, results of qPCR studies on circulating miRNA biomarkers vary greatly and many experiments cannot be reproduced. Missing data in qPCR experiments often occur due to off-target amplification, nonanalyzable qPCR curves and discordance between replicates. The low concentration of most miRNAs leads to most, but not all missing data. Therefore, failure to distinguish between missing data due to a low concentration and missing data due to a randomly occurring technical errors partly explains the variation within and between otherwise similar studies. Based on qPCR kinetics, an analysis pipeline was developed to distinguish missing data due to technical errors from missing data due to a low concentration of the miRNA-equivalent cDNA in the PCR reaction. Furthermore, this pipeline incorporates a method to statistically decide whether concentrations from replicates are sufficiently concordant, which improves stability of results and avoids unnecessary data loss. By going through the pipeline’s steps, the result of each measurement is categorized as “valid, invalid, or undetectable. ” Together with a set of imputation rules, the pipeline leads to more robust and reproducible data as was confirmed experimentally. Using two validation approaches, in two cohorts totaling 2214 heart failure patients, we showed that this pipeline increases both the accuracy and precision of qPCR measurements. In conclusion, this statistical data handling pipeline improves the performance of qPCR studies on low-expressed targets such as circulating miRNAs.
Derechos: Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, sempre que no sigui amb finalitats comercials, i sempre que es reconegui l'autoria de l'obra original. Creative Commons
Lengua: Anglès.
Documento: article ; recerca ; publishedVersion
Materia: Microrna ; Qpcr ; Data analysis
Publicado en: RNA, Vol. 23 Núm. 5 (2017) , p. 811-821

DOI: 10.1261/rna.059063.116
PMID: 28202710

11 p, 713.2 KB

El registro aparece en las colecciones:
Documentos de investigación > Documentos de los grupos de investigación de la UAB > Centros y grupos de investigación (producción científica) > Ciencias de la salud y biociencias > Institut d'Investigació en Ciencies de la Salut Germans Trias i Pujol (IGTP)
Artículos > Artículos de investigación
Artículos > Artículos publicados

 Registro creado el 2018-10-05, última modificación el 2019-03-26

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