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.
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.
Gibellini
Davide
D
Department of Clinical and Experimental Medicine, Microbiology Section, University of Bologna, St. Orsola Hospital, Via Massarenti 9, 40138, Bologna, Italy. davide.gibelline@unibo.it
Vitone
Francesca
F
Gori
Elisa
E
La Placa
Michele
M
Re
Maria Carla
MC
D023241:Antiretroviral Therapy, Highly Active; D001483:Base Sequence; D018791:CD4 Lymphocyte Count; D017931:DNA Primers / Q000235:genetics; D005456:Fluorescent Dyes; D018023:HIV Seronegativity; D006679:HIV Seropositivity / Q000188:drug therapy / Q000276:immunology / Q000821:virology; D015497:HIV-1 / Q000235:genetics* / Q000302:isolation & purification*; D006801:Humans; D012367:RNA, Viral / Q000097:blood / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012680:Sensitivity and Specificity; D014766:Viremia / Q000821:virology; D014773:Virology / Q000379:methods* / Q000706:statistics & numerical data
D017931:DNA Primers; D005456:Fluorescent Dyes; D012367:RNA, Viral