Quantitation of genomic DNA in plasma and serum samples: higher concentrations of genomic DNA found in serum than in plasma.

Author(s): Lee TH, Montalvo L, Chrebtow V, Busch MP

Publication: Transfusion, 2001, Vol. 41, Page 276-82

PubMed ID: 11239235 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 anticoagulant type, blood component, and refrigerated storage on the yield of cell-free DNA from plasma and serum. Aliquots of plasma and serum were removed from the original collection tube at each time-point and were frozen at -80 degrees C until DNA extraction. Cell-free DNA was quantified by PCR amplification of the cell surface recepor HLA DQ alpha and the Y chromosome along with standards followed by radiographic analysis.

   Summary of Findings:

   While only 2 of 18 ACD plasma specimens refrigerated for 2 days contained cell-free DNA, all 18 of the matched 2 day old serum specimens contained cell-free DNA. Fresh serum had more than 20-fold higher concentrations of cell-free DNA than fresh ACD or EDTA plasma. During refrigerated storage, cell-free DNA increased rapidly in serum reaching a 3-fold increase over initial levels observed by day 1 and a 42-fold increase observed between days 1 and 4. In contrast, cell-free DNA concentrations in plasma increased 30-fold between day 0 and 4 of refrigerated storage. When buffy coat from males was added to non-coagulated female blood, the resultant serum contained male DNA indicating lysis of red blood cells during coagulation, but male DNA was not detected in plasma until day 4 of storage.

   Biospecimens

   • Bodily Fluid - Plasma
   • Bodily Fluid - Serum
   • Bodily Fluid - Buffy Coat
   • Bodily Fluid - Blood

   Preservative Types

   • Frozen

   Diagnoses:

   • Normal

   Platform:

   Analyte	Technology Platform
   DNA	Southern blot
   DNA	PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Aliquots and Components	Blood and blood products	Buffy coat Plasma Serum
   Storage	Storage duration	2 h 1 day 2 days 3 days 4 days 5 days 6 days 7 days
   Southern blot Specific	Targeted nucleic acid	HLA DQ alpha Sex-determining region Y
   Biospecimen Acquisition	Anticoagulant	Acid-citrate-dextrose EDTA None

   View more

2. Study Purpose

   The purpose of this study was to determine the effect of DNA isolation kit type on the yield of cell-free DNA using plasma from human immunodeficiency virus (HIV) positive patients. Aliquots of plasma were frozen at -80 degrees C until DNA extraction.

   Summary of Findings:

   The authors report the highest yield of cell-free plasma DNA with the GlasPac/GS and HIV Monitor Assay protocols, and they also report that the consistency of amplification was highest using the HIV Monitor Assay.

   Biospecimens

   • Bodily Fluid - Plasma

   Preservative Types

   • Frozen

   Diagnoses:

   • AIDS/HIV-related

   Platform:

   Analyte	Technology Platform
   DNA	PCR
   DNA	Southern blot

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Analyte Extraction and Purification	Analyte isolation method	GlasPac GeneClean Dynal beads QIAamp blood HIV monitor assay

   View more