Table of Contents
Biotechnology Research International
Volume 2011, Article ID 964831, 10 pages
http://dx.doi.org/10.4061/2011/964831
Research Article

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

1Molecular Biology Department, Centro de InmunoEnsayo (CIE), Calle 134 y Avenida 25 Playa, Apartado Postal 6653, Ciudad de la Habana, CP 11600, Cuba
2HCV Department, Vaccine Division, Centro de Ingeniería Genética y Biotecnología (CIGB), Apartado Postal 6162, Ciudad de la Habana, CP 10600, Cuba

Received 10 January 2011; Revised 19 April 2011; Accepted 27 April 2011

Academic Editor: Gabriel A. Monteiro

Copyright © 2011 Anny Armas Cayarga et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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  log10 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 . 9 2 5 ) 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 = 1 4 ). 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.