title: Generation of HIV-1 and Internal Control Transcripts as Standards for an In-House Quantitative Competitive RT-PCR Assay to Determine HIV-1 Viral Load.
abstract: Human immunodeficiency virus type-1 (HIV-1) viral load is useful for monitoring disease progression in HIV-infected individuals. We generated RNA standards of HIV-1 and internal control (IC) by in vitro transcription and evaluated its performance in a quantitative reverse transcription polymerase chain reaction (qRT-PCR) assay. HIV-1 and IC standards were obtained at high RNA concentrations, without DNA contamination. When these transcripts were included as standards in a qRT-PCR assay, it was obtained a good accuracy (±0.5 log(10) unit of the expected results) in the quantification of the HIV-1 RNA international standard and controls. The lower limit detection achieved using these standards was 511.0 IU/mL. A high correlation (r = 0.925) was obtained between the in-house qRT-PCR assay and the NucliSens easyQ HIV-1 test (bioMerieux) for HIV-1 RNA quantitation with clinical samples (N = 14). HIV-1 and IC RNA transcripts, generated in this study, proved to be useful as standards in an in-house qRT-PCR assay for determination of HIV-1 viral load.
authors.affiliation: Molecular Biology Department, Centro de InmunoEnsayo (CIE), Calle 134 y Avenida 25 Playa, Apartado Postal 6653, Ciudad de la Habana, CP 11600, Cuba.
authors.forename: Anny
authors.initials: A
authors.lastname: Armas Cayarga
authors.forename: Yenitse
authors.initials: Y
authors.lastname: Perea Hernández
authors.forename: Yaimé J
authors.initials: YJ
authors.lastname: González González
authors.forename: Santiago
authors.initials: S
authors.lastname: Dueñas Carrera
authors.forename: Idania
authors.initials: I
authors.lastname: González Pérez
authors.forename: René
authors.initials: R
authors.lastname: Robaina Álvarez