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The performance of human cytomegalovirus digital PCR reference measurement procedure in seven external quality assessment schemes over four years.

Authors
  • Milavec, Mojca1
  • Pavšič, Jernej2
  • Bogožalec Košir, Alexandra2
  • Jones, Gerwyn M3
  • O'Sullivan, Denise M3
  • Devonshire, Alison S3
  • Van Heuverswyn, Fran4
  • Karczmarczyk, Maria4
  • Neeb, Jannika5
  • Plauth, Annabell5
  • Corbisier, Philippe4
  • Schimmel, Heinz4
  • Kummrow, Andreas5
  • Neukammer, Jörg5
  • Foy, Carole A3
  • Kammel, Martin6
  • Grunert, Hans-Peter7
  • Zeichhardt, Heinz8
  • Huggett, Jim F9
  • 1 Department of Biotechnology and Systems Biology, National Institute of Biology, Večna pot 111, 1000 Ljubljana, Slovenia. Electronic address: [email protected] , (Slovenia)
  • 2 Department of Biotechnology and Systems Biology, National Institute of Biology, Večna pot 111, 1000 Ljubljana, Slovenia. , (Slovenia)
  • 3 National Measurement Laboratory (NML), LGC, Queens Road, Teddington, Middlesex TW11 0LY, United Kingdom. , (United Kingdom)
  • 4 European Commission, Joint Research Centre (JRC), Geel, Belgium. , (Belgium)
  • 5 Physikalisch Technische Bundesanstalt, Abbestr. 2-12, D-10587 Berlin, Germany. , (Germany)
  • 6 INSTAND, Gesellschaft zur Foerderung der Qualitaetssicherung in medizinischen Laboratorien e.V., Ubierstr.20, D-40223 Düsseldorf, Germany; IQVD GmbH, Institut fuer Qualitaetssicherung in der Virusdiagnostik, Potsdamer Chaussee 80, D-14129 Berlin, Germany. , (Germany)
  • 7 GBD Gesellschaft fuer Biotechnologische Diagnostik mbH, Berlin, Potsdamer Chaussee 80, D-14129 Berlin, Germany. , (Germany)
  • 8 INSTAND, Gesellschaft zur Foerderung der Qualitaetssicherung in medizinischen Laboratorien e.V., Ubierstr.20, D-40223 Düsseldorf, Germany; IQVD GmbH, Institut fuer Qualitaetssicherung in der Virusdiagnostik, Potsdamer Chaussee 80, D-14129 Berlin, Germany; GBD Gesellschaft fuer Biotechnologische Diagnostik mbH, Berlin, Potsdamer Chaussee 80, D-14129 Berlin, Germany. , (Germany)
  • 9 National Measurement Laboratory (NML), LGC, Queens Road, Teddington, Middlesex TW11 0LY, United Kingdom; School of Biosciences & Medicine, Faculty of Health & Medical Science, University of Surrey, Guildford, United Kingdom. , (United Kingdom)
Type
Published Article
Journal
Methods
Publisher
Elsevier
Publication Date
May 01, 2022
Volume
201
Pages
65–73
Identifiers
DOI: 10.1016/j.ymeth.2021.03.016
PMID: 33812016
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

A candidate digital PCR (dPCR)-based reference measurement procedure for quantification of human cytomegalovirus (hCMV) was evaluated in 10 viral load comparison schemes (seven external quality assessment (EQA) and three additional training schemes) organized by INSTAND e.V. over four years (between September 2014 and March 2018). Four metrology institutes participated in these schemes using the same extraction method and dPCR measurement procedure for the hCMV specific target sequence of UL54 gene. The calibration independent reference measurement procedure results from the metrology institutes were compared to the results of the clinical diagnostic laboratories applying hCMV qPCR measurement procedures calibrated to reference materials. While the criteria for the acceptable deviation from the target value interval for INSTAND's EQA schemes is from -0.8 log10 to +0.8 log10, the majority of dPCR results were between -0.2 log10 to +0.2 log10. Only 4 out of 45 results exceeded this interval with the maximum deviation of -0.542 log10. In the training schemes containing samples with lower hCMV concentrations, more than half of the results deviated less than ±0.2 log10 from the target value, while more than 95% deviated less than ±0.4 log10 from the target value. Evaluation of intra- and inter-laboratory variation of dPCR results confirmed high reproducibility and trueness of the method. This work demonstrates that dPCR has the potential to act as a calibration independent reference measurement procedure for the value assignment of hCMV calibration and reference materials to support qPCR calibration as well as ultimately for routine hCMV load testing. Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.

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