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Workshop report on the extraction of foetal DNA from maternal plasma

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Author

  • Tobias J. Legler
  • Zhong Liu
  • Ariadni Mavrou
  • Kirstin Finning
  • Ilona Hromadnikova
  • Silvia Galbiati
  • Cathy Meaney
  • Maj A. Hulten
  • Francesco Crea
  • Martin L Olsson
  • Deborah G. Maddocks
  • Dorothy Huang
  • Sylvia Armstrong Fisher
  • Markus Sprenger-Haussels
  • Aicha Ait Soussan
  • C. Ellen van der Schoot
Show all

Summary, in English

Objective Cell free foetal DNA (cff DNA) extracted from maternal plasma is now recognized as a potential source for prenatal diagnosis but the methodology is currently not well standardized. To evaluate different manual and automated DNA extraction methods with a view to developing standards, an International Workshop was performed. Methods Three plasma pools from RhD-negative pregnant women, a DNA standard, real-time-PCR protocol, primers and probes for RHD were sent to 12 laboratories and also to one company (Qiagen, Hilden, Germany). In pre-tests, pool 3 showed a low cff DNA concentration, pool I showed a higher concentration and pool 2 an intermediate concentration. Results The QIAamp DSP Virus Kit, the High Pure PCR Template Preparation Kit, an in-house protocol using the QIAamp DNA Blood Mini Kit, the CST genomic DNA purification kit, the Magna Pure LC, the MDx, the M48, the EZl and an in-house protocol using magnetic beads for manual and automated extraction were the methods that were able to reliably detect foetal RHD. The best results were obtained with the QIAamp DSP Virus Kit. The QIAamp DNA Blood Mini Kit showed very comparable results in laboratories that followed the manufacturer's protocol and started with >= 500 mu L plasma. One participant using the QIAamp DNA Blood Midi Kit failed to detect reliably RHD in pool 3. Conclusions This workshop initiated a standardization process for extraction of cff DNA in maternal plasma. The highest yield was obtained by the QIAamp DSP Virus Kit, a result that will be evaluated in more detail in future studies. Copyright (c) 2007 John Wiley & Sons, Ltd.

Publishing year

2007

Language

English

Pages

824-829

Publication/Series

Prenatal Diagnosis

Volume

27

Issue

9

Document type

Journal article

Publisher

John Wiley & Sons Inc.

Topic

  • Obstetrics, Gynecology and Reproductive Medicine

Keywords

  • cell-free foetal DNA
  • non-invasive prenatal diagnosis
  • maternal plasma
  • genotyping
  • RhD
  • sensitivity

Status

Published

Research group

  • Transfusion Medicine

ISBN/ISSN/Other

  • ISSN: 1097-0223