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Impact of two different commercial DNA extraction methods on BK virus viral load

Massimiliano Bergallo, Ilaria Galliano, Elisa Loiacono, Francesca Ferro, Paola Montanari, Paolo Ravanini
  • Massimiliano Bergallo
    Department of Public Health and Paediatric Sciences, University of Turin Medical School, Turin; Laboratory of Citoimmunodiagnostics, University Hospital of City Science and Health, Regina Margherita Children's Hospital, Turin, Italy | massimiliano.bergallo@unito.it
  • Ilaria Galliano
    Department of Public Health and Paediatric Sciences, University of Turin Medical School, Turin; Laboratory of Citoimmunodiagnostics, University Hospital of City Science and Health, Regina Margherita Children's Hospital, Turin, Italy
  • Elisa Loiacono
    Nephrology, Dialysis and Transplantation, University Hospital City of Science and Health, Regina Margherita Children's Hospital, Turin, Italy
  • Francesca Ferro
    Laboratory of Citoimmunodiagnostics, University Hospital of City Science and Health, Regina Margherita Children's Hospital, Turin, Italy
  • Paola Montanari
    Department of Public Health and Paediatric Sciences, University of Turin Medical School, Turin; Laboratory of Citoimmunodiagnostics, University Hospital of City Science and Health, Regina Margherita Children's Hospital, Turin, Italy
  • Paolo Ravanini
    Laboratory of Molecular Virology, Azienda Ospedaliero-Universitaria Maggiore della Carità, Novara, Italy

Abstract

Background and aim: BK virus, a member of human polyomavirus family, is a worldwide distributed virus characterized by a seroprevalence rate of 70-90% in adult population. Monitoring of viral replication is made by evaluation of BK DNA by quantitative polymerase chain reaction. Many different methods can be applied for extraction of nucleic acid from several specimens. The aim of this study was to assess the impact of two different DNA extraction procedure on BK viral load.
Materials and methods: DNA extraction procedure including the Nuclisens easyMAG platform (bioMerieux, Marcy l’Etoile, France) and manual QIAGEN extraction (QIAGEN Hilden, Germany). BK DNA quantification was performed by Real Time TaqMan PCR using a commercial kit.
Result and discussion: The samples capacity, cost and time spent were compared for both systems. In conclusion our results demonstrate that automated nucleic acid extraction method using Nuclisense easyMAG was superior to manual protocol (QIAGEN Blood Mini kit), for the extraction of BK virus from serum and urine specimens.

Keywords

DNA extraction, BKV viral load, PCR real time, transplant recipients

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Submitted: 2014-11-11 13:59:46
Published: 2016-03-31 00:00:00
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Copyright (c) 2016 Massimiliano Bergallo, Ilaria Galliano, Elisa Loiacono, Francesca Ferro, Paola Montanari, Paolo Ravanini

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