title: Assessment of a molecular diagnostic platform for integrated isolation and quantification of mRNA in whole blood.
abstract: Implementation of point-of-care tests may facilitate the health management of infectious diseases by reducing the timeframe on pathogen identification and host response measurements, allowing for immediate diagnosis and guided clinical intervention. In this feasibility study, a novel totally integrated and fully automated real-time polymerase chain reaction (PCR) platform (Idylla™, Biocartis) was assessed to determine the mRNA expression levels of multiple genes from 1 mL of whole blood. To this purpose, a sample-in result-out assay, including mRNA extraction and RT-qPCR-based detection, was ported to the platform. The genes used (matrix metallopeptidase 9, olfactomedin 4, NB1 glycoprotein and lipocalin 2) were previously identified as predictive for severity of disease caused by infection with respiratory syncytial virus (RSV). The reproducibility and robustness of the prototype assay was determined using the blood samples of 21 healthy donors. The data showed that the Idylla™ platform allows for a fast and user-friendly determination of the relative expression levels of the four selected mRNA markers.
authors: Department of Pediatrics, Laboratory of Pediatric Infectious Diseases, Radboud University Medical Center, Nijmegen, The Netherlands. corne.vandenkieboom@radboudumc.nl.|Radboud Institute for Molecular Life Sciences, Nijmegen, The Netherlands. corne.vandenkieboom@radboudumc.nl.
authors: C H
authors: CH
authors: van den Kieboom
authors: Department of Pediatrics, Laboratory of Pediatric Infectious Diseases, Radboud University Medical Center, Nijmegen, The Netherlands.|Radboud Institute for Molecular Life Sciences, Nijmegen, The Netherlands.
authors: G
authors: G
authors: Ferwerda
authors: R&D Department, Biocartis N.V., Mechelen, Belgium.
authors: I
authors: I
authors: de Baere
authors: R&D Department, Biocartis N.V., Mechelen, Belgium.
authors: H
authors: H
authors: Vermeiren
authors: Department of Pediatrics, Laboratory of Pediatric Infectious Diseases, Radboud University Medical Center, Nijmegen, The Netherlands.
authors: R
authors: R
authors: de Groot
authors: R&D Department, Biocartis N.V., Mechelen, Belgium.
authors: R
authors: R
authors: Rossau
authors: Department of Pediatrics, Laboratory of Pediatric Infectious Diseases, Radboud University Medical Center, Nijmegen, The Netherlands.|Radboud Institute for Molecular Life Sciences, Nijmegen, The Netherlands.
authors: M I
authors: MI
authors: de Jonge
mesh_terms: D000328:Adult; D057205:Automation, Laboratory / Q000379:methods; D001774:Blood Chemical Analysis / Q000379:methods*; D005240:Feasibility Studies; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D008297:Male; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D018357:Respiratory Syncytial Virus Infections / Q000175:diagnosis* / Q000473:pathology; D013997:Time Factors; D055815:Young Adult
chemical_list: D012333:RNA, Messenger