title: Workshop report on the extraction of foetal DNA from maternal plasma.
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 > or = 500 microL 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.
authors.affiliation: Department of Transfusion Medicine, University of Göttingen, Göttingen, Germany. tlegler@med.uni-goettingen.de
authors.forename: Tobias J
authors.initials: TJ
authors.lastname: Legler
authors.forename: Zhong
authors.initials: Z
authors.lastname: Liu
authors.forename: Ariadni
authors.initials: A
authors.lastname: Mavrou
authors.forename: Kirstin
authors.initials: K
authors.lastname: Finning
authors.forename: Ilona
authors.initials: I
authors.lastname: Hromadnikova
authors.forename: Silvia
authors.initials: S
authors.lastname: Galbiati
authors.forename: Cathy
authors.initials: C
authors.lastname: Meaney
authors.forename: Maj A
authors.initials: MA
authors.lastname: Hultén
authors.forename: Francesco
authors.initials: F
authors.lastname: Crea
authors.forename: Martin L
authors.initials: ML
authors.lastname: Olsson
authors.forename: Deborah G
authors.initials: DG
authors.lastname: Maddocks
authors.forename: Dorothy
authors.initials: D
authors.lastname: Huang
authors.forename: Sylvia Armstrong
authors.initials: SA
authors.lastname: Fisher
authors.forename: Markus
authors.initials: M
authors.lastname: Sprenger-Haussels
authors.forename: Aicha Ait
authors.initials: AA
authors.lastname: Soussan
authors.forename: C Ellen
authors.initials: CE
authors.lastname: van der Schoot
mesh_terms: D001331:Automation; D004247:DNA / Q000097:blood* / Q000302:isolation & purification*; D005260:Female; D005333:Fetus / Q000378:metabolism; D005820:Genetic Testing / Q000379:methods* / Q000592:standards; D006801:Humans; D033261:Maternal-Fetal Relations; D009035:Mothers; D016133:Polymerase Chain Reaction / Q000379:methods; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods / Q000592:standards
chemical_list: D004247:DNA