Quantitative detection of human immunodeficiency virus type 1 (HIV-1) viral load by SYBR green real-time RT-PCR technique in HIV-1 seropositive patients

J Virol Methods. 2004 Feb;115(2):183-9. doi: 10.1016/j.jviromet.2003.09.030.


HIV-1 viral load represents a basic marker for evaluation of the rate and severity of HIV-1 related disease and to monitor the effectiveness of treatment. An SYBR green-based real-time RT-PCR (SYBR green real-time RT-PCR) revealed by Light Cycler technology was evaluated for quantitation of HIV-1 RNA viral load in plasma of HIV-1 seropositive patients. The performance of the SYBR green real-time PCR was assessed on 56 HIV-1 seropositive patients under highly active retroviral therapy (HAART) and 25 blood donors. The results demonstrated that this technique detected 50 HIV-1 RNA copies per millilitre of plasma. Moreover, we compared real-time RT-PCR with the b-DNA technique considered widely a reference technique for HIV-1 RNA viral load measurement. The parallel quantitative analysis of HIV-1 positive samples showed a high correlation (r=0.908) between the two methods. Although b-DNA and the real-time-based method gave similar sensitivity, the assay determined quantitatively HIV-1 RNA copies in 4 out of 16 samples shown as undetectable by b-DNA. The SYBR green real-time RT-PCR represents a good alternative to b-DNA assay in HIV-1 viral load determination especially during the monitoring of HAART treatment.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antiretroviral Therapy, Highly Active
  • Base Sequence
  • CD4 Lymphocyte Count
  • DNA Primers / genetics
  • Fluorescent Dyes
  • HIV Seronegativity
  • HIV Seropositivity / drug therapy
  • HIV Seropositivity / immunology
  • HIV Seropositivity / virology
  • HIV-1 / genetics*
  • HIV-1 / isolation & purification*
  • Humans
  • RNA, Viral / blood
  • RNA, Viral / genetics
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Reverse Transcriptase Polymerase Chain Reaction / statistics & numerical data
  • Sensitivity and Specificity
  • Viremia / virology
  • Virology / methods*
  • Virology / statistics & numerical data


  • DNA Primers
  • Fluorescent Dyes
  • RNA, Viral