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.
Enviado por Sharilyn Almodovar el
Título | 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. |
Publication Type | Journal Article |
Year of Publication | 2006 |
Autores | Contreras-Galindo, R, González, M, Almodóvar, S, González-Ramírez, S, Lorenzo, E, Yamamura, Y |
Journal | J Virol Methods |
Volume | 136 |
Issue | 1-2 |
Pagination | 51-7 |
Date Published | 2006 Sep |
ISSN | 0166-0934 |
Palabras clave | Analysis of Variance, Antiretroviral Therapy, Highly Active, Base Sequence, Cloning, Molecular, Endogenous Retroviruses, Fluorescence, HIV Infections, HIV-1, Humans, Molecular Sequence Data, Organic Chemicals, Reproducibility of Results, Reverse Transcriptase Polymerase Chain Reaction, RNA, Viral, Sensitivity and Specificity, Staining and Labeling, Viral Load |
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. |
DOI | 10.1016/j.jviromet.2006.03.029 |
Alternate Journal | J. Virol. Methods |
PubMed ID | 16678919 |