Selection of the optimal manual method of cell free fetal DNA isolation from maternal plasma.
Author(s): Repiská G, Sedlackova, Szemes T, Celec P, Minárik G
Publication: Clin Chem Lab Med, 2013, Vol. 51, Page 1185-9
PubMed ID: 23241606 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
The purpose of this study was to compare the cell-free DNA yield between three different Qiagen DNA extraction kits. Blood from pregnant women was stored at 4-8 degrees C for less than 24 h before separation of plasma by double centrifugation. The plasma was stored frozen at -20 degrees C until extraction. Cell-free DNA was quantified by amplification of the DYS14 marker in the Testis specific protein Y linked gene (TSPY), and total circulating DNA was quantified by amplification of the X-chromosome specific androgen receptor gene region.
Summary of Findings:
Significantly more cell-free fetal DNA was obtained from plasma using the DSP virus kit or the CNA kit than the DBM kit (p<0.001 and p<0.0001, respectively), but there was no difference in cell-free fetal DNA yield between the DSP virus kit and the CNA kit. When the input volume and elution volume for the DBM kit were adjusted to those used by the DSP kit (500 uL and 50 uL, respectively), significantly more cell-free fetal DNA was still extracted using the DSP kit than the DBM kit (p<0.0001), but there was no difference in total circulating DNA yield.
Biospecimens
Preservative Types
- Frozen
Diagnoses:
- Pregnant
Platform:
Analyte Technology Platform DNA Real-time qPCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Aliquots and Components Aliquot size/volume 200 uL
500 uL
1 mL
Analyte Extraction and Purification Analyte isolation method DNA blood mini kit
DSP virus kit
Circulating nucleic acid kit
Analyte Extraction and Purification Rehydration of dried sample/specimen 50 uL
100 uL
200 uL
Real-time qPCR Specific Nucleic acid amplification DYS14
X-chromosome specific androgen receptor