Effects of specimen collection, processing, and storage conditions on stability of human immunodeficiency virus type 1 RNA levels in plasma.
To define the optimal blood collection parameters for plasma human immunodeficiency virus type 1 (HIV-1) viral load testing, plasma HIV-1 RNA levels were quantitated with the NASBA HIV-1 RNA QT System from blood specimens that were collected, processed, and stored under a variety of conditions that might have affected HIV-1 RNA stability. We determined that when whole blood was processed within 2 h of specimen collection the levels of HIV-1 RNA detected in EDTA-, heparin-, and acid citrate dextrose (ACD)-anticoagulated plasma samples were comparable. The levels of HIV-1 RNA in serum specimens (mean = 4.126 log units) were significantly lower (P < 0.01) than the levels in corresponding plasma samples (mean = 4.501 log units). One cycle of freeze-thaw (-70 degrees C) did not significantly reduce the level of HIV-1 RNA detected in EDTA-, heparin-, or ACD-anticoagulated plasmas. The EDTA-anticoagulated plasmas showed the smallest decrease in HIV-1 RNA copies (0.050 log units). HIV-1 RNA levels decreased over a 6-month time period in serum as well as in EDTA-, ACD-, and heparin-anticoagulated plasmas stored at -70 degrees C. However, the only significant decreases were for serum (mean decrease = 0.317 log units) and heparin-anticoagulated samples (mean decrease = 0.384 log units). A comparison of the levels of HIV-1 RNA in cell-free plasma collected in VACUTAINER EDTA Plasma Preparation Tubes and in standard VACUTAINER EDTA tubes determined that HIV-1 RNA levels were stable for up to 30 h after collection when stored at either room temperature (mean standard deviation [SD] = +/- 0.101 log units) or at 4 degrees C (mean SD = +/- 0.102 log units) as cell-free plasma or as EDTA-anticoagulated whole blood (mean SD = +/- 0.109 log units). These data indicate that EDTA-anticoagulated plasma is the most suitable and stable matrix for HIV-1 RNA quantitation.
Ginocchio
C C
CC
Department of Medicine, North Shore University Hospital, Manhasset, New York 11030, USA. cginocch@nshs.edu
Wang
X P
XP
Kaplan
M H
MH
Mulligan
G
G
Witt
D
D
Romano
J W
JW
Cronin
M
M
Carroll
R
R
D000925:Anticoagulants; D001793:Blood Preservation / Q000379:methods; D001800:Blood Specimen Collection / Q000295:instrumentation / Q000379:methods*; D019343:Citric Acid; D015925:Cryopreservation / Q000379:methods; D004355:Drug Stability; D004492:Edetic Acid; D005947:Glucose / Q000031:analogs & derivatives; D006679:HIV Seropositivity / Q000097:blood* / Q000821:virology; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006493:Heparin; D006801:Humans; D007202:Indicators and Reagents; D010949:Plasma; D012367:RNA, Viral / Q000097:blood*; D015203:Reproducibility of Results; D019562:Viral Load / Q000295:instrumentation / Q000379:methods
D000925:Anticoagulants; D007202:Indicators and Reagents; D012367:RNA, Viral; C002113:acid citrate dextrose; D019343:Citric Acid; D006493:Heparin; D004492:Edetic Acid; D005947:Glucose