Workshop report on the extraction of foetal DNA from maternal plasma

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

Research output: Contribution to journalArticle

Abstract

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 1 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 EZ1 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 μL 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.

Original languageEnglish
Pages (from-to)824-829
Number of pages6
JournalPrenatal Diagnosis
Volume27
Issue number9
DOIs
Publication statusPublished - Sep 2007

Keywords

  • Cell-free foetal DNA
  • Genotyping
  • Maternal plasma
  • Non-invasive prenatal diagnosis
  • RhD
  • Sensitivity

ASJC Scopus subject areas

  • Genetics(clinical)
  • Obstetrics and Gynaecology

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