TY - JOUR
T1 - Consensus guidelines for the validation of qRT-PCR assays in clinical research by the CardioRNA consortium
AU - EU-CardioRNA COST Action CA17129
AU - de Gonzalo-Calvo, David
AU - Marchese, Monica
AU - Hellemans, Jan
AU - Betsou, Fay
AU - Skov Frisk, Nanna Lond
AU - Dalgaard, Louise Torp
AU - Lakkisto, Päivi
AU - Foy, Carole
AU - Scherer, Andreas
AU - Garcia Bermejo, María Laura
AU - Devaux, Yvan
PY - 2022/3/10
Y1 - 2022/3/10
N2 - 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.
AB - 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.
KW - assay validation
KW - clinical research assays
KW - noncoding RNA
KW - PCR
KW - RNA
KW - assay validation
KW - clinical research assays
KW - noncoding RNA
KW - PCR
KW - RNA
U2 - 10.1016/j.omtm.2021.12.007
DO - 10.1016/j.omtm.2021.12.007
M3 - Review
AN - SCOPUS:85123118381
SN - 2329-0501
VL - 24
SP - 171
EP - 180
JO - Molecular Therapy - Methods and Clinical Development
JF - Molecular Therapy - Methods and Clinical Development
ER -