Automated extraction improves multiplex molecular detection of infection in septic patients.

Sepsis is one of the leading causes of morbidity and mortality in hospitalized patients worldwide. Molecular technologies for rapid detection of microorganisms in patients with sepsis have only recently become available. LightCycler SeptiFast test M(grade) (Roche Diagnostics GmbH) is a multiplex PCR...

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Autores principales:	Benito J Regueiro, Eduardo Varela-Ledo, Lucia Martinez-Lamas, Javier Rodriguez-Calviño, Antonio Aguilera, Antonio Santos, Antonio Gomez-Tato, Julian Alvarez-Escudero
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2010
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/eebd59342a0b4daab149116553f97652
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spelling	oai:doaj.org-article:eebd59342a0b4daab149116553f976522021-11-18T07:03:22ZAutomated extraction improves multiplex molecular detection of infection in septic patients.1932-620310.1371/journal.pone.0013387https://doaj.org/article/eebd59342a0b4daab149116553f976522010-10-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/20967222/?tool=EBIhttps://doaj.org/toc/1932-6203Sepsis is one of the leading causes of morbidity and mortality in hospitalized patients worldwide. Molecular technologies for rapid detection of microorganisms in patients with sepsis have only recently become available. LightCycler SeptiFast test M(grade) (Roche Diagnostics GmbH) is a multiplex PCR analysis able to detect DNA of the 25 most frequent pathogens in bloodstream infections. The time and labor saved while avoiding excessive laboratory manipulation is the rationale for selecting the automated MagNA Pure compact nucleic acid isolation kit-I (Roche Applied Science, GmbH) as an alternative to conventional SeptiFast extraction. For the purposes of this study, we evaluate extraction in order to demonstrate the feasibility of automation. Finally, a prospective observational study was done using 106 clinical samples obtained from 76 patients in our ICU. Both extraction methods were used in parallel to test the samples. When molecular detection test results using both manual and automated extraction were compared with the data from blood cultures obtained at the same time, the results show that SeptiFast with the alternative MagNA Pure compact extraction not only shortens the complete workflow to 3.57 hrs., but also increases sensitivity of the molecular assay for detecting infection as defined by positive blood culture confirmation.Benito J RegueiroEduardo Varela-LedoLucia Martinez-LamasJavier Rodriguez-CalviñoAntonio AguileraAntonio SantosAntonio Gomez-TatoJulian Alvarez-EscuderoPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 5, Iss 10, p e13387 (2010)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Benito J Regueiro Eduardo Varela-Ledo Lucia Martinez-Lamas Javier Rodriguez-Calviño Antonio Aguilera Antonio Santos Antonio Gomez-Tato Julian Alvarez-Escudero Automated extraction improves multiplex molecular detection of infection in septic patients.
description	Sepsis is one of the leading causes of morbidity and mortality in hospitalized patients worldwide. Molecular technologies for rapid detection of microorganisms in patients with sepsis have only recently become available. LightCycler SeptiFast test M(grade) (Roche Diagnostics GmbH) is a multiplex PCR analysis able to detect DNA of the 25 most frequent pathogens in bloodstream infections. The time and labor saved while avoiding excessive laboratory manipulation is the rationale for selecting the automated MagNA Pure compact nucleic acid isolation kit-I (Roche Applied Science, GmbH) as an alternative to conventional SeptiFast extraction. For the purposes of this study, we evaluate extraction in order to demonstrate the feasibility of automation. Finally, a prospective observational study was done using 106 clinical samples obtained from 76 patients in our ICU. Both extraction methods were used in parallel to test the samples. When molecular detection test results using both manual and automated extraction were compared with the data from blood cultures obtained at the same time, the results show that SeptiFast with the alternative MagNA Pure compact extraction not only shortens the complete workflow to 3.57 hrs., but also increases sensitivity of the molecular assay for detecting infection as defined by positive blood culture confirmation.
format	article
author	Benito J Regueiro Eduardo Varela-Ledo Lucia Martinez-Lamas Javier Rodriguez-Calviño Antonio Aguilera Antonio Santos Antonio Gomez-Tato Julian Alvarez-Escudero
author_facet	Benito J Regueiro Eduardo Varela-Ledo Lucia Martinez-Lamas Javier Rodriguez-Calviño Antonio Aguilera Antonio Santos Antonio Gomez-Tato Julian Alvarez-Escudero
author_sort	Benito J Regueiro
title	Automated extraction improves multiplex molecular detection of infection in septic patients.
title_short	Automated extraction improves multiplex molecular detection of infection in septic patients.
title_full	Automated extraction improves multiplex molecular detection of infection in septic patients.
title_fullStr	Automated extraction improves multiplex molecular detection of infection in septic patients.
title_full_unstemmed	Automated extraction improves multiplex molecular detection of infection in septic patients.
title_sort	automated extraction improves multiplex molecular detection of infection in septic patients.
publisher	Public Library of Science (PLoS)
publishDate	2010
url	https://doaj.org/article/eebd59342a0b4daab149116553f97652
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Automated extraction improves multiplex molecular detection of infection in septic patients.

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