Influence of temperature during transportation on cell-free DNA analysis.

Author(s): Hidestrand M, Stokowski R, Song K, Oliphant A, Deavers J, Goetsch M, Simpson P, Kuhlman R, Ames M, Mitchell M, Tomita-Mitchell A

Publication: Fetal Diagn Ther, 2012, Vol. 31, Page 122-8

PubMed ID: 22261730 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of transporting whole blood in EDTA or Streck BCT on quantification of total and fetal cf-DNA in plasma from pregnant women carrying male fetuses. For each tube type, one specimen from each patient was immediately processed to plasma and stored at -80 degrees C until DNA extraction. The other tubes were shipped at room temperature (17-25.5 degrees C) or on frozen gel packs (two 3.5-7 degrees C, 1 7.5-20 degrees C), slowly brought to room temperature and processed to plasma 72 h after collection. Plasma from shipped specimens was shipped on dry ice back to the original site where it was stored at -80 degrees C until DNA extraction. Fetal DNA was quantified by TaqMan real-time qPCR amplification of Sex determining region of the Y chromosome (SRY), and total cf-DNA was quantified by TaqMan RNAse P assay.

   Summary of Findings:

   Total cf-DNA levels were comparable in specimens collected in either tube type and processed to plasma immediately or shipped at room temperature as whole blood in BCT tubes before processing to plasma, but total cf-DNA levels were higher when whole blood was shipped in K3 EDTA tubes at room temperature before processing (p<0.001). Further, total cf-DNA levels were lower in specimens processed immediately than when whole blood was shipped on frozen gel packs in BCT tubes (p<0.05) or K3 EDTA tubes (p<0.001). In contrast, the quantity of fetal cf-DNA was not affected by shipment and storage in either BCT or K3 EDTA tubes, regardless of shipping temperature. The percentage of fetal cf-DNA in plasma increased with increasing gestational age in both tube types when blood was processed immediately. However, the percentage of fetal cf-DNA was significantly lower when whole blood was shipped in K3 EDTA tubes than when whole blood was processed to plasma immediately (p<0.001), regardless of shipping temperature.

   Biospecimens

   • Bodily Fluid - Blood
   • Bodily Fluid - Plasma

   Preservative Types

   • None (Fresh)
   • Streck/BCT

   Diagnoses:

   • Pregnant

   Platform:

   Analyte	Technology Platform
   DNA	Real-time qPCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Between site transportation method	Air courier Not transported
   Storage	Specimen transport duration/condition	On ice At room temperature
   Biospecimen Acquisition	Type of collection container/solution	K3 EDTA Vacutainer Streck BCT tube
   Preaquisition	Diagnosis/ patient condition	19-22 weeks pregnant 31-36 weeks pregnant
   Biospecimen Aliquots and Components	Centrifugation	Centrifugation delays investigated
   Biospecimen Preservation	Type of fixation/preservation	None (fresh) Blood collection tube additive

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