Preanalytical stability of HIV-1 and HCV RNA: impact of storage and plasma separation from cells on blood donation testing by NAT.

Author(s): Schulze TJ, Weiss C, Luhm J, Brockmann C, Görg S, Hennig H

Publication: Transfus Med, 2011, Vol. 21, Page 99-106

PubMed ID: 21092012 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 and storage of plasma on HIV and HCV viral load. Whole blood spiked with HCV and HIV was stored for up to 3 days at 5 or 21 degrees C prior to centrifugation. Subsequently, plasma was stored at 5 degrees C with and without cells and assayed daily until day 5.

   Summary of Findings:

   The average HIV CT increased when centrifugation was delayed for up to 72 h, with the most significant increase observed between 18 and 48 h. After whole blood was held for 48 h at 21 degrees C, 3 of 20 spiked specimens were negative for HIV, but only 1 of 14 was negative after 48 h at 5 degrees C. In contrast to HIV, there was no clear decrease in HCV concentration with delays to centrifugation at 5 or 21 degrees C, and only one specimen was found to be HCV negative after delayed centrifugation. HIV levels in stored plasma were not significantly affected by the temperature at which whole blood was kept during delayed centrifugation or the presence of cells during storage, however, there were more HIV detection failures in specimens that were not centrifuged until 48 h after collection than in those centrifuged within 18 or 24 h of collection. There were also more HIV detection failures in plasma obtained after a 24 h delay prior to centrifugation at 5 degrees C and stored as plasma in the presence of cells compared to plasma obtained after the same centrifugation delay but transferred to a new tube and stored without cells. HCV was generally less affected by delayed centrifugation and storage of plasma than HIV, and only 4 of 103 spiked specimens became HCV negative under any experimental conditions compared to 33 of 114 spiked specimens for HIV. The HCV specimens that became negative were all subjected to centrifugation delays at 21 degrees C for at least 48 h and were subsequently stored as plasma for an additional 48 h prior to assay. The authors recommend storage of whole blood at 5 degrees C for 24 h or less when measuring HIV or HCV viral loads in plasma.

   Biospecimens

   • Bodily Fluid - Blood
   • Bodily Fluid - Plasma

   Preservative Types

   • None (Fresh)

   Diagnoses:

   • AIDS/HIV-related
   • Hepatitis
   • Not specified

   Platform:

   Analyte	Technology Platform
   RNA	Real-time qRT-PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Storage duration	0 h 18 h 24 h 48 h 3 days 4 days 5 days
   Storage	Storage conditions	Uncentrifuged blood Plasma in new tube Plasma on cells
   Storage	Storage temperature	5 degrees C 21 degrees C
   Biospecimen Aliquots and Components	Blood and blood products	Plasma Whole blood
   Biospecimen Aliquots and Components	Centrifugation	Centrifugation delays investigated

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