Evaluation of sample stability and automated DNA extraction for fetal sex determination using cell-free fetal DNA in maternal plasma.
Author(s): Ordoñez E, Rueda L, Cañadas MP, Fuster C, Cirigliano V
Publication: Biomed Res Int, 2013, Vol. 2013, Page 195363
PubMed ID: 24222898 PubMed Review Paper? No
Purpose of Paper
The purpose of this paper was to determine the effect of DNA extraction method on the quantification of fetal sex-determining region Y (SRY) in maternal plasma after blood specimens were refrigerated for more than 24 h.
Conclusion of Paper
While the manual DNA extraction method led to lower average SRY cycle threshold (CT) values than the automated method, the cell-free fetal DNA (cffDNA) yields were more variable using the manual method.
Studies
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Study Purpose
The purpose of this study was to determine the effect of DNA extraction method on the quantification of fetal SRY in plasma from maternal blood that had been refrigerated for more than 24 h. Potassium EDTA blood was collected from 158 women during the first trimester of pregnancy (108 carrying a male fetus and 50 carrying a female fetus). After transport of the blood to the laboratory and >24 h refrigerated storage, plasma was obtained through double centrifugation. Plasma was stored at -20°C until DNA extraction.
Summary of Findings:
For plasma obtained after >24 h of refrigerated blood storage, the manual QIAamp DSP virus extraction kit led to lower average SRY CT values than the automated COBAS AmpliPrep DNA/RNA extractor (36.59 versus 37.74). However, the cffDNA yields were more variable using the manual extraction method.
Biospecimens
Preservative Types
- Frozen
Diagnoses:
- Pregnant
Platform:
Analyte Technology Platform DNA Real-time qPCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Real-time qPCR Specific Targeted nucleic acid SRY
Analyte Extraction and Purification Analyte isolation method QIAamp DSP virus kit (Manual)
COBAS AmpliPrep DNA/RNA extractor (Automated)