A multiplex real-time PCR for quantification of HIV-1 DNA and the human albumin gene in CD4+ cells

Authors

  • LARS E. ERIKSSON,

    1. Department of Virology, Swedish Institute for Infectious Disease Control, Solna,
    2. Microbiology and Tumor Biology Center, Karolinska Institutet, Stockholm, and
    3. Department of Nursing, Karolinska Institutet, Huddinge, Sweden
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  • THOMAS LEITNER,

    1. Department of Virology, Swedish Institute for Infectious Disease Control, Solna,
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  • BRITTA WAHREN,

    1. Department of Virology, Swedish Institute for Infectious Disease Control, Solna,
    2. Microbiology and Tumor Biology Center, Karolinska Institutet, Stockholm, and
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  • ANN-CHARLOTTE BOSTRÖM,

    1. Department of Virology, Swedish Institute for Infectious Disease Control, Solna,
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  • KERSTIN I. FALK

    1. Department of Virology, Swedish Institute for Infectious Disease Control, Solna,
    2. Microbiology and Tumor Biology Center, Karolinska Institutet, Stockholm, and
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Lars E. Eriksson, Department of Virology, Swedish Institute for Infectious Disease Control, SE-171 82 Solna, Sweden. e-mail: Lars.Eriksson@omv.ki.se

Abstract

We have established a simple system for measuring HIV-1 DNA load in CD4+ cells. In a multiplex configuration, a conserved region in the HIV-1 pol gene and a section of the human albumin gene were simultaneously amplified to estimate the number of HIV-1 DNA copies per cellular genome. An established Epstein-Barr virus (EBV) standard system was used to calibrate the HIV-1 quantification. Our multiplex PCR system was tested on different in vitro developed HIV-1 strains and on longitudinal samples from eight patients. The system was able to amplify both in vitro and in vivo samples of various genetic compositions. In all eight patients, HIV-1 DNA was detected and ranged between 0.17 and 51×10−3 copies per CD4+ cell and could be monitored longitudinally, including long-term PI-ART and STI. The measured HIV-1 DNA load may be used to select the best time for the institution or re-institution of therapy.

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