title: A new Real-Time-RT-PCR for quantitation of human endogenous retroviruses type K (HERV-K) RNA load in plasma samples: increased HERV-K RNA titers in HIV-1 patients with HAART non-suppressive regimens.
abstract: Viral components of the human endogenous retroviruses type K (HERV-K) have been largely detected in plasma from HIV-1 infected individuals. A Sybr Green Real-Time RT-PCR approach was optimized for detection and quantitation of HERV-K RNA titers in plasma samples using the iCycler technology. The method detected 1000 HERV-K RNA copies/mL of plasma sample. The Intra- and Inter-assay performance revealed a coefficient of variations that ranged from 0.2 to 2.46%, demonstrating accuracy and reproducibility. We quantified the HERV-K RNA load in 20 HIV-1 patients receiving highly active antiretroviral therapy (HAART). We found increased HERV-K RNA titers in patients with non-suppressive HAART (patients who may develop drug-resistance and/or received suboptimal therapeutic doses), compared to suppressive regimens (p < 0.001). HERV-K RNA was not detected in HCV-1 positive or seronegative controls. Sequencing of Real-Time RT-PCR products revealed particular HERV-K subtypes activated in the HIV-1 infection. The application of this assay could expand the understanding of the role of HERV-K in the HIV-1 infection and others pathological conditions.
authors: AIDS Research Program, Ponce School of Medicine, Area Académica 395, Zona Ind. Reparada 2, Ponce, PR 00716-2348, USA. rafcont@hotmail.com
authors: Rafael
authors: R
authors: Contreras-Galindo
authors: Marta
authors: M
authors: González
authors: Sharilyn
authors: S
authors: Almodovar-Camacho
authors: Sandra
authors: S
authors: González-Ramírez
authors: Eric
authors: E
authors: Lorenzo
authors: Yasuhiro
authors: Y
authors: Yamamura