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Workshop report on the extraction of foetal DNA from maternal plasma
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Legler, Tobias J., Liu, Zhong, Mavrou, Ariadni, Finning, Kirstin, Hromadnikova, Ilona, Galbiati, Silvia, Meaney, Cathy, Hultén, Maj A., Crea, Francesco , Olsson, Martin L., Maddocks, Deborah G., Huang, Dorothy, Fisher, Sylvia Armstrong, Sprenger-Haussels, Markus, Soussan, Aicha Ait and van der Schoot, C. Ellen (2007) Workshop report on the extraction of foetal DNA from maternal plasma. Prenatal Diagnosis, Vol.27 (No.9). pp. 824-829. doi:10.1002/pd.1783 ISSN 0197-3851.
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Official URL: http://dx.doi.org/10.1002/pd.1783
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.
| Item Type: | Journal Article | ||||
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| Subjects: | Q Science > QH Natural history > QH426 Genetics R Medicine > RG Gynecology and obstetrics |
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| Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) > Biological Sciences ( -2010) | ||||
| Journal or Publication Title: | Prenatal Diagnosis | ||||
| Publisher: | John Wiley & Sons Ltd. | ||||
| ISSN: | 0197-3851 | ||||
| Official Date: | September 2007 | ||||
| Dates: |
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| Volume: | Vol.27 | ||||
| Number: | No.9 | ||||
| Number of Pages: | 6 | ||||
| Page Range: | pp. 824-829 | ||||
| DOI: | 10.1002/pd.1783 | ||||
| Status: | Peer Reviewed | ||||
| Publication Status: | Published | ||||
| Access rights to Published version: | Restricted or Subscription Access |
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