NIH, National Cancer Institute, Division of Cancer Treatment and Diagnosis (DCTD) NIH - National Institutes of Health National Cancer Institute DCTD - Division of Cancer Treatment and Diagnosis

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

The purpose of this paper was to determine the effects of extraction method on the yield of DNA from blood.

Conclusion of Paper

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 for different length of time, the authors to not discuss the effects of specimen storage on DNA yield or integrity.

Studies

  1. Study Purpose

    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.

    Biospecimens
    Preservative Types
    • Other Preservative
    • Frozen
    Diagnoses:
    • Not specified
    Platform:
    AnalyteTechnology Platform
    DNA Real-time qPCR
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Biospecimen Preservation Type of fixation/preservation Frozen
    Refrigeration
    Analyte Extraction and Purification Analyte isolation method Bead-based
    Column-based
    Precipitation-based

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