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
Conclusion of Paper
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.
- None (Fresh)
Analyte Technology Platform RNA Branch DNA assay RNA RT-PCR
Classification Pre-analytical Factor Value(s) Storage Storage temperature -80 degrees C
-20 degrees C
4 degrees C
Storage Freeze/thaw cycling 0 cycles
Biospecimen Aliquots and Components Blood and blood products Serum
Branch DNA assay Specific Technology platform RT-PCR
Storage Storage duration 0 days
Storage Time at room temperature 0 h
Biospecimen Aliquots and Components Centrifugation Centrifugation delays investigated