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Comparative evaluation of the QIAsymphony RGQ system with the easyMAG/R-gene combination for the quantitation of cytomegalovirus DNA load in whole blood

BACKGROUND: The detection of cytomegalovirus (CMV) DNA in blood is a key feature of the virological surveillance of immunocompromised patients. METHODS: The QIAsymphony RGQ system (QIAGEN S.A.S., France) combines the extraction/distribution steps on QIAsymphony SP/AS instruments with amplification o...

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Autores principales: Pillet, Sylvie, Bourlet, Thomas, Pozzetto, Bruno
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3485198/
https://www.ncbi.nlm.nih.gov/pubmed/23046712
http://dx.doi.org/10.1186/1743-422X-9-231
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author Pillet, Sylvie
Bourlet, Thomas
Pozzetto, Bruno
author_facet Pillet, Sylvie
Bourlet, Thomas
Pozzetto, Bruno
author_sort Pillet, Sylvie
collection PubMed
description BACKGROUND: The detection of cytomegalovirus (CMV) DNA in blood is a key feature of the virological surveillance of immunocompromised patients. METHODS: The QIAsymphony RGQ system (QIAGEN S.A.S., France) combines the extraction/distribution steps on QIAsymphony SP/AS instruments with amplification on a Rotor-Gene Q RT-PCR machine. This system was compared to a strategy combining an extraction step on the NUCLISENS easyMAG platform (bioMérieux) with the CMV R-gene kit (Argene) on 100 whole blood specimens collected from immunocompromised patients of the University Hospital of Saint-Etienne, France. RESULTS: The overall agreement between the two strategies was 86% (kappa coefficient of 0.67); the 14 discrepant results corresponded to low DNA loads. The 62 samples found positive with both tests were correlated (Pearson r coefficient of 0.70, P < 0.01) despite an over quantitation of 0.25 log(10) copies/ml with the easyMAG/Argene strategy (P < 0.001). Very close results were also obtained with a commercial panel of 10 samples with CMV loads ranging from 2.36 to 6.41 log(10) copies/ml. The inter-run and intra-run variability was consistently lower with the QIAGEN platform. CONCLUSIONS: These results validate the performance of the QIAsymphony RGQ system for the routine quantitation of CMV DNA. This fully-automated platform reduces the hands-on time and improves standardization, traceability and quality control assessment.
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spelling pubmed-34851982012-11-01 Comparative evaluation of the QIAsymphony RGQ system with the easyMAG/R-gene combination for the quantitation of cytomegalovirus DNA load in whole blood Pillet, Sylvie Bourlet, Thomas Pozzetto, Bruno Virol J Research BACKGROUND: The detection of cytomegalovirus (CMV) DNA in blood is a key feature of the virological surveillance of immunocompromised patients. METHODS: The QIAsymphony RGQ system (QIAGEN S.A.S., France) combines the extraction/distribution steps on QIAsymphony SP/AS instruments with amplification on a Rotor-Gene Q RT-PCR machine. This system was compared to a strategy combining an extraction step on the NUCLISENS easyMAG platform (bioMérieux) with the CMV R-gene kit (Argene) on 100 whole blood specimens collected from immunocompromised patients of the University Hospital of Saint-Etienne, France. RESULTS: The overall agreement between the two strategies was 86% (kappa coefficient of 0.67); the 14 discrepant results corresponded to low DNA loads. The 62 samples found positive with both tests were correlated (Pearson r coefficient of 0.70, P < 0.01) despite an over quantitation of 0.25 log(10) copies/ml with the easyMAG/Argene strategy (P < 0.001). Very close results were also obtained with a commercial panel of 10 samples with CMV loads ranging from 2.36 to 6.41 log(10) copies/ml. The inter-run and intra-run variability was consistently lower with the QIAGEN platform. CONCLUSIONS: These results validate the performance of the QIAsymphony RGQ system for the routine quantitation of CMV DNA. This fully-automated platform reduces the hands-on time and improves standardization, traceability and quality control assessment. BioMed Central 2012-10-09 /pmc/articles/PMC3485198/ /pubmed/23046712 http://dx.doi.org/10.1186/1743-422X-9-231 Text en Copyright ©2012 Pillet et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Pillet, Sylvie
Bourlet, Thomas
Pozzetto, Bruno
Comparative evaluation of the QIAsymphony RGQ system with the easyMAG/R-gene combination for the quantitation of cytomegalovirus DNA load in whole blood
title Comparative evaluation of the QIAsymphony RGQ system with the easyMAG/R-gene combination for the quantitation of cytomegalovirus DNA load in whole blood
title_full Comparative evaluation of the QIAsymphony RGQ system with the easyMAG/R-gene combination for the quantitation of cytomegalovirus DNA load in whole blood
title_fullStr Comparative evaluation of the QIAsymphony RGQ system with the easyMAG/R-gene combination for the quantitation of cytomegalovirus DNA load in whole blood
title_full_unstemmed Comparative evaluation of the QIAsymphony RGQ system with the easyMAG/R-gene combination for the quantitation of cytomegalovirus DNA load in whole blood
title_short Comparative evaluation of the QIAsymphony RGQ system with the easyMAG/R-gene combination for the quantitation of cytomegalovirus DNA load in whole blood
title_sort comparative evaluation of the qiasymphony rgq system with the easymag/r-gene combination for the quantitation of cytomegalovirus dna load in whole blood
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3485198/
https://www.ncbi.nlm.nih.gov/pubmed/23046712
http://dx.doi.org/10.1186/1743-422X-9-231
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