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

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

   • Bodily Fluid - Blood
   • Bodily Fluid - Serum

   Preservative Types

   • None (Fresh)

   Diagnoses:

   • Hepatitis

   Platform:

   Analyte	Technology Platform
   RNA	Branch DNA assay
   RNA	RT-PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(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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