Advanced

Workshop report on the extraction of foetal DNA from maternal plasma

Legler, Tobias J.; Liu, Zhong; Mavrou, Ariadni; Finning, Kirstin; Hromadnikova, Ilona; Galbiati, Silvia; Meaney, Cathy; Hulten, Maj A.; Crea, Francesco and Olsson, Martin L LU , et al. (2007) In Prenatal Diagnosis 27(9). p.824-829
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 I showed a higher concentration and pool 2 an intermediate concentration. Results The QIAamp DSP Virus Kit, the High Pure PCR Template... (More)
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. (Less)
Please use this url to cite or link to this publication:
author
, et al. (More)
(Less)
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
cell-free foetal DNA, non-invasive prenatal diagnosis, maternal plasma, genotyping, RhD, sensitivity
in
Prenatal Diagnosis
volume
27
issue
9
pages
824 - 829
publisher
John Wiley and Sons Ltd
external identifiers
  • wos:000249713000006
  • scopus:34548757303
ISSN
1097-0223
DOI
10.1002/pd.1783
language
English
LU publication?
yes
id
98cef689-9203-411a-a7d0-aedb042b16da (old id 655880)
date added to LUP
2007-12-14 13:52:59
date last changed
2017-10-22 03:35:06
@article{98cef689-9203-411a-a7d0-aedb042b16da,
  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 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.},
  author       = {Legler, Tobias J. and Liu, Zhong and Mavrou, Ariadni and Finning, Kirstin and Hromadnikova, Ilona and Galbiati, Silvia and Meaney, Cathy and Hulten, Maj A. and Crea, Francesco and Olsson, Martin L and Maddocks, Deborah G. and Huang, Dorothy and Fisher, Sylvia Armstrong and Sprenger-Haussels, Markus and Soussan, Aicha Ait and van der Schoot, C. Ellen},
  issn         = {1097-0223},
  keyword      = {cell-free foetal DNA,non-invasive prenatal diagnosis,maternal plasma,genotyping,RhD,sensitivity},
  language     = {eng},
  number       = {9},
  pages        = {824--829},
  publisher    = {John Wiley and Sons Ltd},
  series       = {Prenatal Diagnosis},
  title        = {Workshop report on the extraction of foetal DNA from maternal plasma},
  url          = {http://dx.doi.org/10.1002/pd.1783},
  volume       = {27},
  year         = {2007},
}