Literature DB >> 22296792

A new highly automated extraction system for quantitative real-time PCRs from whole blood samples: routine monitoring of opportunistic infections in immunosuppressed patients.

C Mengelle1, J-M Mansuy, K Sauné, C Barthe, J Boineau, J Izopet.   

Abstract

BACKGROUND: Rapid, high throughput extraction systems are needed to monitor viral infections in immunosuppressed patients.
OBJECTIVES: Evaluate the performance of the MagNA Pure 96™ extraction system, and compare it to the COBAS Ampliprep™ for quantitative real-time PCR from whole blood samples. STUDY
DESIGN: Compare the MagNA Pure LC™, COBAS Ampliprep™ and MagNA Pure 96™ using ten-fold dilutions of blood samples containing cytomegalovirus. Evaluate analytical performances of the MagNA Pure 96™ from test samples containing cytomegalovirus. Evaluate clinical performances from 209 blood samples collected prospectively, extracted with the COBAS Ampliprep™ and the MagNA Pure 96™ systems and tested for cytomegalovirus, Epstein-Barr, BK and JC viruses.
RESULTS: All three extraction systems gave similar results with dilutions of a cytomegalovirus-positive sample. Analytical tests showed that the limit of detection was 500 copies/ml, specificity was 100%, with no cross-contamination. Quantification was linear from 3.0 to 6.0 log(10)copies/ml. Intra-assay variation was 8.3-0.9% and inter-assay variation 8.8-5.2%. Clinical specimens extracted with the MagNA Pure 96™ and COBAS Ampliprep™ instruments agreed well for cytomegalovirus (r=0.54; p=0.07), Epstein-Barr virus (0.69; p=0.0005) and BK virus (0.85; p=0.01). All 55 samples were negative for JC virus. Mean loads were similar for cytomegalovirus (0.17 log(10)copies/ml) and BK virus (-0.24 log(10)copies/ml) while that of Epstein-Barr virus was slightly lower (1.02 log(10)copies/ml).
CONCLUSIONS: The MagNA Pure 96™ instrument is an easy-to-use, reliable high throughput platform for extracting nucleic acid from clinical whole blood specimens. Copyright Â
© 2012 Elsevier B.V. All rights reserved.

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Year:  2012        PMID: 22296792     DOI: 10.1016/j.jcv.2012.01.002

Source DB:  PubMed          Journal:  J Clin Virol        ISSN: 1386-6532            Impact factor:   3.168


  2 in total

1.  Fully automated quantification of cytomegalovirus (CMV) in whole blood with the new sensitive Abbott RealTime CMV assay in the era of the CMV international standard.

Authors:  Nathalie Schnepf; Catherine Scieux; Matthieu Resche-Riggon; Linda Feghoul; Alienor Xhaard; Sébastien Gallien; Jean-Michel Molina; Gérard Socié; Denis Viglietti; François Simon; Marie-Christine Mazeron; Jérôme Legoff
Journal:  J Clin Microbiol       Date:  2013-04-24       Impact factor: 5.948

2.  Performance Evaluation of the Real-Q Cytomegalovirus (CMV) Quantification Kit Using Two Real-Time PCR Systems for Quantifying CMV DNA in Whole Blood.

Authors:  Jong Eun Park; Ji Youn Kim; Sun Ae Yun; Myoung Keun Lee; Hee Jae Huh; Jong Won Kim; Chang Seok Ki
Journal:  Ann Lab Med       Date:  2016-11       Impact factor: 3.464

  2 in total

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