Lars E Eriksson | PubMed Health

PubMed2003

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

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 51x10-3 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.

Base Sequence↗

PubMed2003

HIV type 1 DNA development during long-term supervised therapy interruption.

We studied the pattern of HIV-1 DNA development and the association to other HIV-related factors during long-term supervised therapy interruption (LT-STI). Fifteen patients were treated with long-time protease inhibitor-based antiretroviral therapy (PI-ART). They had HIV-1 RNA at <50 copies/ml over 33.4 (SD 9.5) months and CD4(+) cell counts of 875 (SD 415) x 10(6)/liter. A real-time polymerase chain reaction, amplifying fragments of the HIV-1 pol gene and the human albumin gene simultaneously, was used to quantify HIV-1 DNA molecules in CD4(+) cells. The quantity of HIV-1 DNA in CD4(+) cells increased during LT-STI in all 15 patients, with an average doubling time of 2 months. Tentatively, three patterns were observed: rapid initial increase with subsequent stabilizing levels, rapid continuous increase, and slow increase. The HIV-1 DNA slope was positively related to the HIV-1 RNA maximum and steady state level and the baseline HIV-1 DNA value. It was inversely related to the decrease in CD4(+) cells both before the start of PI-ART and during the LT-STI. To conclude, HIV-1 DNA persists in infected CD4(+) cells despite long-term effective PI-ART and will increase after therapy interruption. The most important clinical predictor of long-term STI failure was the rapid CD4(+) cell decline before PI-ART. In patients with a steep pre-PI-ART slope it may be prudent to continue treatment and not initiate therapy interruption.

Anti-HIV Agents↗