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Assessment of a molecular diagnostic platform for integrated isolation and quantification of mRNA in whole blood
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 integra...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Berlin Heidelberg
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4607718/ https://www.ncbi.nlm.nih.gov/pubmed/26298058 http://dx.doi.org/10.1007/s10096-015-2470-2 |
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author | van den Kieboom, C. H. Ferwerda, G. de Baere, I. Vermeiren, H. de Groot, R. Rossau, R. de Jonge, M. I. |
author_facet | van den Kieboom, C. H. Ferwerda, G. de Baere, I. Vermeiren, H. de Groot, R. Rossau, R. de Jonge, M. I. |
author_sort | van den Kieboom, C. H. |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-4607718 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-46077182015-10-20 Assessment of a molecular diagnostic platform for integrated isolation and quantification of mRNA in whole blood van den Kieboom, C. H. Ferwerda, G. de Baere, I. Vermeiren, H. de Groot, R. Rossau, R. de Jonge, M. I. Eur J Clin Microbiol Infect Dis Original Article 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. Springer Berlin Heidelberg 2015-08-23 2015 /pmc/articles/PMC4607718/ /pubmed/26298058 http://dx.doi.org/10.1007/s10096-015-2470-2 Text en © The Author(s) 2015 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. |
spellingShingle | Original Article van den Kieboom, C. H. Ferwerda, G. de Baere, I. Vermeiren, H. de Groot, R. Rossau, R. de Jonge, M. I. Assessment of a molecular diagnostic platform for integrated isolation and quantification of mRNA in whole blood |
title | Assessment of a molecular diagnostic platform for integrated isolation and quantification of mRNA in whole blood |
title_full | Assessment of a molecular diagnostic platform for integrated isolation and quantification of mRNA in whole blood |
title_fullStr | Assessment of a molecular diagnostic platform for integrated isolation and quantification of mRNA in whole blood |
title_full_unstemmed | Assessment of a molecular diagnostic platform for integrated isolation and quantification of mRNA in whole blood |
title_short | Assessment of a molecular diagnostic platform for integrated isolation and quantification of mRNA in whole blood |
title_sort | assessment of a molecular diagnostic platform for integrated isolation and quantification of mrna in whole blood |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4607718/ https://www.ncbi.nlm.nih.gov/pubmed/26298058 http://dx.doi.org/10.1007/s10096-015-2470-2 |
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