LTR real-time PCR for HIV-1 DNA quantitation in blood cells for early diagnosis in infants born to seropositive mothers treated in HAART area (ANRS CO 01)

J Med Virol. 2009 Feb;81(2):217-23. doi: 10.1002/jmv.21390.


HIV-1 diagnosis in babies born to seropositive mothers is one of the challenges of HIV epidemics in children. A simple, rapid protocol was developed for quantifying HIV-1 DNA in whole blood samples and was used in the ANRS French pediatric cohort in conditions of prevention of mother-to-child transmission. A quantitative HIV-1 DNA protocol (LTR real-time PCR) requiring small blood volumes was developed. First, analytical reproducibility was evaluated on 172 samples. Results obtained on blood cell pellets and Ficoll-Hypaque separated mononuclear cells were compared in 48 adult HIV-1 samples. Second, the protocol was applied to HIV-1 diagnosis in infants in parallel with plasma HIV-RNA quantitation. This prospective study was performed in children born between May 2005 and April 2007 included in the ANRS cohort. The assay showed good reproducibility. The 95% detection cut-off value was 6 copies/PCR, that is, 40 copies/10(6) leukocytes. HIV-DNA levels in whole blood were highly correlated with those obtained after Ficoll-Hypaque separation (r = 0.900, P < 0.0001). A total of 3,002 specimens from 1,135 infants were tested. The specificity of HIV-DNA and HIV-RNA assays was 100%. HIV-1 infection was diagnosed in nine infants before age 60 days. HIV-DNA levels were low, underlining the need for sensitive assays when highly active antiretroviral therapy (HAART) has been given. The performances of this HIV-DNA assay showed that it is adapted to early diagnosis in children. The results were equivalent to those of HIV-RNA assay. HIV-DNA may be used even in masked primary infection in newborns whose mothers have received HAART.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Anti-HIV Agents / therapeutic use
  • Antiretroviral Therapy, Highly Active*
  • DNA, Viral / analysis
  • DNA, Viral / blood*
  • DNA, Viral / genetics
  • Female
  • HIV Infections / diagnosis*
  • HIV Infections / drug therapy
  • HIV Infections / immunology
  • HIV Infections / transmission
  • HIV Seropositivity*
  • HIV-1 / genetics
  • HIV-1 / immunology
  • HIV-1 / isolation & purification*
  • Humans
  • Infant
  • Infant, Newborn
  • Leukocytes, Mononuclear / virology
  • Mothers
  • Polymerase Chain Reaction / methods*
  • Pregnancy
  • Prospective Studies
  • Reproducibility of Results
  • Sensitivity and Specificity


  • Anti-HIV Agents
  • DNA, Viral