This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Kirstein, L. M.
Right arrow Articles by Muñoz, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kirstein, L. M.
Right arrow Articles by Muñoz, A.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, August 1999, p. 2428-2433, Vol. 37, No. 8
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Effects of Anticoagulant, Processing Delay, and Assay Method (Branched DNA versus Reverse Transcriptase PCR) on Measurement of Human Immunodeficiency Virus Type 1 RNA Levels in Plasma

Lynn M. Kirstein,1,* John W. Mellors,2 Charles R. Rinaldo Jr.,2 Joseph B. Margolick,1 Janis V. Giorgi,3 John P. Phair,4 Edith Dietz,1 Phalguni Gupta,2 Christopher H. Sherlock,5 Robert Hogg,5 J. S. G. Montaner,5 and Alvaro Muñoz1

Departments of Epidemiology and Molecular Microbiology and Immunology, Johns Hopkins School of Hygiene and Public Health, Baltimore, Maryland1; University of Pittsburgh School of Medicine, Pittsburgh Veterans Affairs Medical Center, and Graduate School of Public Health, Pittsburgh, Pennsylvania2; Department of Medicine, University of California at Los Angeles School of Medicine, Los Angeles, California3; Northwestern University Medical School, Chicago, Illinois4; and University of British Columbia, Vancouver, British Columbia, Canada5

Received 25 February 1999/Returned for modification 31 March 1999/Accepted 29 April 1999

We conducted two studies to determine the potential influence of delays in blood processing, type of anticoagulant, and assay method on human immunodeficiency virus type 1 (HIV-1) RNA levels in plasma. The first was an experimental study in which heparin- and EDTA-anticoagulated blood samples were collected from 101 HIV-positive individuals and processed to plasma after delays of 2, 6, and 18 h. HIV-1 RNA levels in each sample were then measured by both branched-DNA (bDNA) and reverse transcriptase PCR (RT-PCR) assays. Compared to samples processed within 2 h, the loss (decay) of HIV-1 RNA in heparinized blood was significant (P < 0.05) but small after 6 h (bDNA assay, -0.12 log10 copies/ml; RT-PCR, -0.05 log10 copies/ml) and after 18 h (bDNA assay, -0.27 log10 copies/ml; RT-PCR, -0.15 log10 copies/ml). Decay in EDTA-anticoagulated blood was not significant after 6 h (bDNA assay, -0.002 log10 copies/ml; RT-PCR, -0.02 log10 copies/ml), but it was after 18 h (bDNA assay, -0.09 log10 copies/ml; RT-PCR, -0.09 log10 copies/ml). Only 4% of samples processed after 6 h lost more than 50% (>= 0.3 log10 copies/ml) of the HIV-1 RNA, regardless of the anticoagulant or the assay that was used. The second study compared HIV-1 RNA levels in samples from the Multicenter AIDS Cohort Study (MACS; samples were collected in heparin-containing tubes in 1985, had a 6-h average processing delay, and were assayed by bDNA assay) and the British Columbia Drug Treatment Program (BCDTP) (collected in EDTA- or acid citrate dextrose-containing tubes in 1996 and 1997, had a 2-h maximum processing delay, and were assayed by RT-PCR). HIV-1 RNA levels in samples from the two cohorts were not significantly different after adjusting for CD4+-cell count and converting bDNA assay values to those corresponding to the RT-PCR results. In summary, the decay of HIV-1 RNA measured in heparinized blood after 6 h was small (-0.05 to -0.12 log10 copies/ml), and the minor impact of this decay on HIV-1 RNA concentrations in archived plasma samples of the MACS was confirmed by the similarity of CD4+-cell counts and assay-adjusted HIV-1 RNA concentrations in the MACS and BCDTP.

* Corresponding author. Mailing address: Department of Epidemiology, Johns Hopkins School of Public Health, 615 N. Wolfe St., Room E7007, Baltimore, MD 21205. Phone: (410) 614-1340. Fax: (410) 614-7125. E-mail: lkirstei{at}jhsph.edu.

Journal of Clinical Microbiology, August 1999, p. 2428-2433, Vol. 37, No. 8
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Stewart, M., Desport, M., Hartaningsih, N., Wilcox, G. (2005). TaqMan Real-Time Reverse Transcription-PCR and JDVp26 Antigen Capture Enzyme-Linked Immunosorbent Assay To Quantify Jembrana Disease Virus Load during the Acute Phase of In Vivo Infection. J. Clin. Microbiol. 43: 5574-5580 [Abstract] [Full Text]  
  • Ahdieh-Grant, L., Yamashita, T. E., Phair, J. P., Detels, R., Wolinsky, S. M., Margolick, J. B., Rinaldo, C. R., Jacobson, L. P. (2003). When to Initiate Highly Active Antiretroviral Therapy: A Cohort Approach. Am J Epidemiol 157: 738-746 [Abstract] [Full Text]  
  • Jacobson, L. P., Li, R., Phair, J., Margolick, J. B., Rinaldo, C. R., Detels, R., Munoz, A. (2002). Evaluation of the Effectiveness of Highly Active Antiretroviral Therapy in Persons with Human Immunodeficiency Virus using Biomarker-based Equivalence of Disease Progression. Am J Epidemiol 155: 760-770 [Abstract] [Full Text]  
  • Garcia, M. E., Blanco, J. L., Caballero, J., Gargallo-Viola, D. (2002). Anticoagulants Interfere with PCR Used To Diagnose Invasive Aspergillosis. J. Clin. Microbiol. 40: 1567-1568 [Full Text]  
  • Tarwater, P. M., Mellors, J., Gore, M. E., Margolick, J. B., Phair, J., Detels, R., Munoz, A. (2001). Methods to Assess Population Effectiveness of Therapies in Human Immunodeficiency Virus Incident and Prevalent Cohorts. Am J Epidemiol 154: 675-681 [Abstract] [Full Text]  
  • Jennings, C., Bremer, J. W., Brambilla, D. J. (2000). Investigation of Effects of Acid Citrate Dextrose and EDTA on Ability To Quantitatively Culture Human Immunodeficiency Virus. J. Clin. Microbiol. 38: 3522-3522 [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»