title: Preanalytical stability of HIV-1 and HCV RNA: impact of storage and plasma separation from cells on blood donation testing by NAT.
abstract: BACKGROUND
  The aim of this study was to evaluate the optimal preanalytical conditions prior to nucleic acid amplification technology (NAT) for human immunodeficiency virus-1 (HIV-1) or Hepatitis C virus (HCV) RNA in pools of 96 plasma specimens with regard to storage temperature, time and plasma separation in a blood donation environment.
  
  
  METHODS
  Changes in viral nucleic acid concentration of HIV-1 and HCV were observed for 5 days according to the Paul-Ehrlich-Institute's (PEI) guidelines that demand 95%-detection limit of at least 10 000 IU mL(-1) for HIV-1 RNA and 5000 IU mL(-1) for HCV RNA within a single donor blood specimen. Ninety-five per cent detection limits of HIV-1 RNA over 3 days after storage at either 5 or 21 °C were evaluated by using standardised HIV-1 RNA-positive plasma.
  
  
  RESULTS
  HCV RNA in whole blood samples proved to be more stable than HIV-1 RNA. Whole blood storage at 21 °C was shown to decrease the detectability of HIV-1 RNA even after only 18 h. Plasma samples once used for NAT at time 18 h did not alter viral stability up to 48 h after donation. Ninety-five per cent detection limits of HIV-1 RNA were securely below 10 000 IU mL(-1) for 24 h after whole blood storage at 5 °C.
  
  
  CONCLUSIONS
  These results may lead to a discussion around the most suitable preanalytical conditions in blood donation environments. Contrary to the current PEI guidelines that allow storage of whole blood specimens up to 18 h at 21 °C, these results suggest that immediate storage in a 5 °C container after blood donation is more suitable and would permit storage of whole blood up to 24 h prior to the separation of plasma from cells.
authors.affiliation: Institute of Transfusion Medicine and Immunlogie, German Red Cross Blood Service, Baden Württemberg-Hessen, Medical Faculty of Mannheim, Heidelberg University, Mannheim, Germany.
authors.forename: T J
authors.initials: TJ
authors.lastname: Schulze
authors.forename: C
authors.initials: C
authors.lastname: Weiss
authors.forename: J
authors.initials: J
authors.lastname: Luhm
authors.forename: C
authors.initials: C
authors.lastname: Brockmann
authors.forename: S
authors.initials: S
authors.lastname: Görg
authors.forename: H
authors.initials: H
authors.lastname: Hennig
mesh_terms: D001782:Blood Donors; D001793:Blood Preservation / Q000191:economics / Q000295:instrumentation / Q000379:methods*; D058731:Blood Safety / Q000191:economics / Q000592:standards; D015497:HIV-1 / Q000235:genetics*; D016174:Hepacivirus / Q000235:genetics*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000191:economics; D009994:Osmolar Concentration; D010949:Plasma / Q000737:chemistry; D017410:Practice Guidelines as Topic; D020871:RNA Stability; D012367:RNA, Viral / Q000097:blood*; D012680:Sensitivity and Specificity; D013696:Temperature; D013997:Time Factors; D014186:Transportation
chemical_list: D012367:RNA, Viral