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

Impact of various handling and storage conditions on quantitative detection of hepatitis C virus RNA.

Author(s): Halfon P, Khiri H, Gerolami V, Bourliere M, Feryn JM, Reynier P, Gauthier A, Cartouzou G

Publication: J Hepatol, 1996, Vol. 25, Page 307-11

PubMed ID: 8895009 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to determine the effects of The purpose of this paper was to determine the effects of delayed centrifugation, storage, and freeze-thaw cycling on the quantification of hepatitis C virus (HCV) load in serum.

Conclusion of Paper

HCV load quantification was not affected by storage of serum for 5 days at 4 degrees C or -20 degrees C, or for 6 months at -80 degrees C, but room temperature storage of serum for 5 days resulted in undetectable HCV load. Delayed centrifugation (4 h at room temperature) or 5 freeze-thaw cycles resulted in a significant loss of HCV load compared to levels in immediately analyzed specimens.

Studies

  1. Study Purpose

    The purpose of this study was to determine the effects of delayed centrifugation, storage, and freeze-thaw cycling on the quantification of HCV load in serum. When insufficient RNA was present for quantification via the branched DNA assay, RT-PCR was used.

    Summary of Findings:

    Storage of serum at 4 or -20 degrees C for up to 5 days, or -80 degrees C for 6 months, only resulted in insignificant declines in HCV load compared to immediately analyzed specimens. However, when serum was stored at room temperature for 5 days, or 4 degrees C for 6 months, HCV was undetectable in all 7 specimens. Further, when whole blood was kept for 4 h at room temperature prior to centrifugation and storage of serum at -80 degrees C for 5 days, or when specimens underwent 5 freeze-thaw cycles during the 5 days at -80 degrees C, there was a significant 29.5% and 16% decrease in HCV load, respectively compared to levels in immediately analyzed specimens. Two of the 16 serum specimens that were negative for HCV RNA using the branched DNA assay were positive by RT-PCR.

    Biospecimens
    Preservative Types
    • None (Fresh)
    Diagnoses:
    • Hepatitis
    Platform:
    AnalyteTechnology Platform
    RNA Branch DNA assay
    RNA RT-PCR
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Storage Storage temperature -80 degrees C
    -20 degrees C
    4 degrees C
    Room temperature
    Storage Freeze/thaw cycling 0 cycles
    5 cycles
    Biospecimen Aliquots and Components Blood and blood products Serum
    Whole blood
    Branch DNA assay Specific Technology platform RT-PCR
    Storage Storage duration 0 days
    5 days
    6 months
    Storage Time at room temperature 0 h
    4 h
    Biospecimen Aliquots and Components Centrifugation Centrifugation delays investigated

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