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Journal of Clinical Microbiology, July 2009, p. 2170-2174, Vol. 47, No. 7
0095-1137/09/$08.00+0     doi:10.1128/JCM.00519-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Overestimation of Human Immunodeficiency Virus Type 1 Load Caused by the Presence of Cells in Plasma from Plasma Preparation Tubes{triangledown}

Anne-Marte Bakken Kran,1,3* Tom Øystein Jonassen,1,3 Mette Sannes,2,3 Kirsti Jakobsen,1,3 Andreas Lind,2,3 Arild Mæland,2,3 and Mona Holberg-Petersen1,3

Department of Microbiology, National Reference Laboratory for HIV, Oslo University Hospital, Ullevål, Oslo NO-0407, Norway,1 Department of Infectious Diseases, Oslo University Hospital, Ullevål, Oslo NO-0407, Norway,2 University of Oslo Faculty Division, Oslo, Norway3

Received 13 March 2009/ Returned for modification 17 April 2009/ Accepted 29 April 2009

The human immunodeficiency virus type 1 (HIV-1) load is an important marker of disease progression and treatment efficacy in patients with HIV-1 infection. In recent years, an increase in the number of samples with detectable HIV-1 RNA has been reported among patients with previously suppressed viral loads, affecting clinical patient care and leading to repeat measurements of viral load and drug resistance. This rise seems to have coincided with the increased use of plasma preparation tubes (PPTs) for sample collection, and we have aimed to explain why PPTs might yield elevated HIV-1 RNA levels. The impacts of different sample-processing procedures on HIV-1 RNA levels were compared retrospectively. Prospectively, the presence of different cells and cell-associated HIV-1 nucleic acids in paired plasma samples from PPTs centrifuged before (PPT1) and after (PPT2) transportation to the laboratory was compared. A retrospective analysis of 4,049 patient samples with <1,000 HIV-1 RNA copies/ml showed elevated HIV-1 RNA levels in plasma from PPT1 compared with the levels from PPT2 and standard EDTA-containing tubes. Prospective data revealed cell-associated HIV-1 nucleic acids and abundant blood cells in plasma from PPT1 but not from the corresponding PPT2. The levels of HIV-1 RNA correlated with the lymphocyte counts in plasma in PPT1. Cells could be removed by the recentrifugation of PPT1 before analysis. In conclusion, the transportation of PPTs after centrifugation may render cells in the plasma fraction containing cell-associated HIV-1 nucleic acids that contribute significantly to the HIV-1 RNA copy numbers in patients with low viral loads.

* Corresponding author. Mailing address: National Reference Laboratory for HIV, Department of Microbiology, Oslo University Hospital, Ullevål, Oslo NO-0407, Norway. Phone: 47 22118820. Fax: 47 22406056. E-mail: a.m.b.kran{at}medisin.uio.no

{triangledown} Published ahead of print on 6 May 2009.

Journal of Clinical Microbiology, July 2009, p. 2170-2174, Vol. 47, No. 7
0095-1137/09/$08.00+0     doi:10.1128/JCM.00519-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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