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

• Published in: Journal of virological methods 2004-02, Vol.115 (2), p.183-189
• Main Authors: Gibellini, Davide, Vitone, Francesca, Gori, Elisa, Placa, Michele La, Re, Maria Carla
• Format: Article
• Language: English
• Subjects: Antiretroviral Therapy, Highly Active; Base Sequence; Biological and medical sciences; CD4 Lymphocyte Count; DNA Primers - genetics; Fluorescent Dyes; Fundamental and applied biological sciences. Psychology; HIV Seronegativity; HIV Seropositivity - drug therapy; HIV Seropositivity - immunology; HIV Seropositivity - virology; HIV-1 - genetics; HIV-1 - isolation & purification; HIV-1 viral load; Human immunodeficiency virus 1; Humans; Microbiology; Real-time RT-PCR; Reverse Transcriptase Polymerase Chain Reaction - methods; Reverse Transcriptase Polymerase Chain Reaction - statistics & numerical data; RNA, Viral - blood; RNA, Viral - genetics; Sensitivity and Specificity; SYBR green; Techniques used in virology; Viremia - virology; Virology; Virology - methods; Virology - statistics & numerical data
• ISSN: 0166-0934; 1879-0984
• 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.