Real-time PCR evaluation of cell-free DNA subjected to various storage and shipping conditions.
Author(s): Wang Q, Cai Y, Brady P, Vermeesch JR
Publication: Genet Mol Res, 2015, Vol. 14, Page 12797-804
PubMed ID: 26505430 PubMed Review Paper? No
Purpose of Paper
This paper investigated the effects of tube type on the stability of total and fetal cell-free DNA during shipping for 2 or 5 days at three different temperatures.
Conclusion of Paper
Total and fetal cell-free DNA levels were comparable to that in immediately processed blood when blood was shipped at 10˚C or less in BCT tubes, but significant decreases in the levels of fetal cell-free DNA and increases in total cell-free DNA were noted when blood was shipped in K3EDTA tubes. The changes observed were smaller when blood was shipped at -20 to 0˚C than at 0 to 10˚C. The authors report a decrease in cell-free fetal DNA and an increase in total cell free DNA when shipped at 0 to 30˚C in either K3EDTA or BCT tubes.
Studies
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Study Purpose
This study investigated the effects of tube type on the stability of total and fetal cell-free DNA during shipping for 2 or 5 days at three different temperatures. Duplicate blood specimens from pregnant women carrying male fetuses were collected into each of the tube types (K3EDTA and BCT tubes). To study the effect of transportation, specimens from 100 women were collected into two tubes of each type; one was analyzed immediately and the second was shipped for 2 or 5 days in a styrofoam box and exposed to temperatures of 0°C to 10°, 0°C to 30°C, or -20°C to 0°C. Plasma was obtained by centrifugation of the blood at 2000 x g for 10 min followed by centrifugation at 14000 x g for an unspecified duration. DNA was extracted from plasma using the QIAamp Circulating Nucleic Acid (CNA) kit. DNA was quantified by real-time PCR amplification of β-actin and SRY.
Summary of Findings:
Shipping the blood at 0 to 10˚C for 2 or 5 days in BCT tubes or 2 days in K3EDTA tubes had no impact on the amount of total or fetal cell-free DNA, but fetal cell-free DNA levels were significantly lower (P<0.01) and total DNA levels significantly higher (P<0.01) when blood was shipped for 5 days at 0 to 10˚C in K3EDTA tubes than when blood was processed immediately after collection. Smaller decreases in fetal cell-free DNA levels and increases in total cell-free DNA were noted when specimens were shipped at -20 to 0˚C in K3EDTA tubes than when shipped at 0 to 10˚C, but the authors report a decrease in cell-free fetal DNA and an increase in total cell free DNA in both tube types when shipped at 0 to 30˚C.
Biospecimens
Preservative Types
- None (Fresh)
Diagnoses:
- Pregnant
Platform:
Analyte Technology Platform DNA Real-time qPCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Acquisition Type of collection container/solution Streck BCT tube
K3EDTA tube
Real-time qPCR Specific Targeted nucleic acid β-actin
SRY
Storage Specimen transport duration/condition -20° to 0°C
0° to 10°C
0° to 30°C
2 days
5 days
Storage Between site transportation method Mailed