title: Stability of HIV-1 RNA in blood during specimen handling and storage prior to amplification by NASBA-QT.
abstract: The influence of different storage temperatures and anticoagulation conditions on the HIV-1 RNA load as measured by NASBA-QT was examined. Blood specimens from 14 HIV-1 infected individuals were processed within 2 h after collection. The HIV-1 RNA load remained stable for at least 6 months when samples were frozen directly at -70 degrees C in lysis buffer. This lysis buffer fully inactivated the virus. When whole EDTA blood was stored, the HIV-1 RNA load was stable for 72 h at 25 degrees C, but it declined within 24 h at 4 degrees C. The HIV-1 RNA load in whole heparinized blood declined significantly after 24 h at both 4 and 25 degrees C. It was slightly lower (average of 0.18 log ml-1) than in whole EDTA blood. At 4 degrees C, the HIV-RNA load in serum and EDTA-plasma stored with lysis buffer did not decline up to 14 days. At + 30 degrees C, however, the load declined significantly after 2 days. Of clinical significance, the mean load in EDTA plasma was 0.5 log ml-1 higher than in serum. This difference was patient dependent (range 0.1-0.7 log ml-1). We thus recommend, for quantifying HIV-1 RNA by NASBA, to use preferably EDTA blood which is kept at room temperature until plasma separation. When using heparinized blood, the plasma should be stored frozen within 8 h.
authors.affiliation: Central Laboratory, Netherlands Red Cross Blood Transfusion Service, Department of Viral Diagnostics, Amsterdam.
authors.forename: S M
authors.initials: SM
authors.lastname: Bruisten
authors.forename: P
authors.initials: P
authors.lastname: Oudshoorn
authors.forename: P
authors.initials: P
authors.lastname: van Swieten
authors.forename: B
authors.initials: B
authors.lastname: Boeser-Nunnink
authors.forename: P
authors.initials: P
authors.lastname: van Aarle
authors.forename: S P
authors.initials: SP
authors.lastname: Tondreau
authors.forename: H T
authors.initials: HT
authors.lastname: Cuypers
mesh_terms: D001793:Blood Preservation; D002021:Buffers; D004492:Edetic Acid; D005784:Gene Amplification; D015658:HIV Infections / Q000175:diagnosis / Q000235:genetics* / Q000821:virology; D015497:HIV-1 / Q000235:genetics*; D006493:Heparin / Q000097:blood; D006801:Humans; D006863:Hydrogen-Ion Concentration; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000097:blood* / Q000302:isolation & purification; D011933:Reagent Kits, Diagnostic; D012260:Ribonucleases / Q000037:antagonists & inhibitors; D013048:Specimen Handling; D013696:Temperature
chemical_list: D002021:Buffers; D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic; D006493:Heparin; D004492:Edetic Acid; D012260:Ribonucleases