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© Research
Publication : Molecular therapy. Methods & clinical development

Consensus guidelines for the validation of qRT-PCR assays in clinical research by the CardioRNA consortium.

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Molecular therapy. Methods & clinical development - 10 Mar 2022

de Gonzalo-Calvo D, Marchese M, Hellemans J, Betsou F, Skov Frisk NL, Dalgaard LT, Lakkisto P, Foy C, Scherer A, Garcia Bermejo ML, Devaux Y, ,

Link to Pubmed [PMID] – 35118162

Link to DOI – 10.1016/j.omtm.2021.12.007

Mol Ther Methods Clin Dev 2022 Mar; 24(): 171-180

Despite promising findings, quantitative PCR (qPCR)-based tests for RNA quantification have experienced serious limitations in their clinical application. The noticeable lack of technical standardization remains a huge obstacle in the translation of qPCR-based tests. The incorporation of qPCR-based tests into the clinic will benefit from guidelines for clinical research assay validation. This will ultimately impact the clinical management of the patient, including diagnosis, prognosis, prediction, monitoring of the therapeutic response, and evaluation of toxicity. However, clear assay validation protocols for biomarker investigation in clinical trials using molecular assays are currently lacking. Here, we will focus on the necessary steps, including sample acquisition, processing and storage, RNA purification, target selection, assay design, and experimental design, that need to be taken toward the appropriate validation of qRT-PCR assays in clinical research. These recommendations can fill the gap between research use only (RUO) and in vitro diagnostics (IVD). Our contribution provides a tool for basic and clinical research for the development of validated assays in the intermediate steps of biomarker research. These guidelines are based on the current understanding and consensus within the EU-CardioRNA COST Action consortium (www.cardiorna.eu). Their applicability encompasses all clinical areas.

https://pubmed.ncbi.nlm.nih.gov/35118162