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.
Eriksson
Lars E
LE
Department of Virology, Swedish Institute for Infectious Disease Control, Solna, Sweden. Lars.Eriksson@omv.ki.se
Leitner
Thomas
T
Wahren
Britta
B
Bostrom
Aaa-Charlotte
AC
Falk
Kerstin I
KI
D001483:Base Sequence; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000502:physiology* / Q000821:virology; D004279:DNA, Viral / Q000235:genetics; D015751:Genes, pol / Q000235:genetics; D015658:HIV Infections / Q000097:blood / Q000175:diagnosis / Q000821:virology*; D015497:HIV-1 / Q000235:genetics* / Q000276:immunology; D006801:Humans; D008137:Longitudinal Studies; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA; D012709:Serum Albumin / Q000235:genetics*; D019562:Viral Load
D004279:DNA, Viral; D012367:RNA, Viral; D012709:Serum Albumin