SPIDIA-DNA: An External Quality Assessment for the pre-analytical phase of blood samples used for DNA-based analyses.
Author(s): Malentacchi F, Pazzagli M, Simi L, Orlando C, Wyrich R, Hartmann CC, Verderio P, Pizzamiglio S, Ciniselli CM, Tichopad A, Kubista M, Gelmini S
Publication: Clin Chim Acta, 2013, Vol. 424, Page 274-86
PubMed ID: 23714327 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
The purpose of this study was to determine the effects of extraction method on the yield of DNA from blood as determined by TaqMan real-time PCR. Blood collected into citrate-phosphate-dextrose-adenine (CPDA) was aliquoted into 2 mL polypropylene tubes. Aliquoted CPDA blood was shipped on ice packs and transported internationally throughout Europe by courier. Each laboratory extracted DNA using their own protocol and different storage conditions for blood specimens prior to extraction, including storage ranging from <24 h to >72 h at -80 to 4 degrees C. DNA extracted in the laboratories was returned to the blood collection facility on ice packs and stored at -20 degrees C until analysis.
Summary of Findings:
The average DNA yield, as assessed by TaqMan real-time PCR amplification of the RNaseP gene, was lower when DNA was extracted using a precipitation-based method than a bead based (p=0.002) or column-based (p<0.01) method. Despite analyzing DNA which had been extracted from blood stored at different temperatures and shipped and stored for different length of time, the authors to not discuss the effects of refrigerated or frozen storage of blood on DNA yield or integrity.
- Other Preservative
- Not specified
Analyte Technology Platform DNA Real-time qPCR
Classification Pre-analytical Factor Value(s) Biospecimen Preservation Type of fixation/preservation Frozen
Analyte Extraction and Purification Analyte isolation method Bead-based