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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
biochemistry, genetics and molecular biology
Fluorogenic LUX primer for quantitation of HIV-1 by real-time RT-PCR
Molecular Biotechnology, Volume 32, No. 2, Year 2006
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Description
Measurement of HIV-1 viral load in plasma is an important marker of disease progression and efficacy of antiretroviral therapy. Real-time polymerase chain reaction (PCR) offers an opportunity to develop more affordable alternative viral load assays. This article reports on the development of a novel real-time reverse-transcriptase (RT)-PCR assay for quantitation of HIV-1 RNA copies. This assay utilizes the LightCycler® (version2) real-time PCR platform and light upon extension (LUX) primer for specific detection of amplicons. An external standard (ES) for quantitation of viral RNA represents an in vitro transcribed RNA. The LUX assay shows a wide linear (R2 = 0.99) dynamic range from 4 × 106 to 4 × 102 copies/mL. Analytical sensitivity of the assay is 4 × 102 copies/mL of ES RNA. Intra- and inter-assay variability of the LUX assay was less than 0.5log10 copies of ES RNA (i.e., no clinically significant variability was found). Virology quality assurance (VQA) HIV-1 RNA copy controls were used to validate ES and preliminarily evaluate the assay performance. This feasibility study demonstrated that the LUX assay is sensitive, reproducible, and compares well to the Roche Amplicor tests used for characterization of the RNA copy controls. These results suggest further evaluation of the LUX assay using a large cohort of well-characterized samples from HIV-1 positive individuals. © 2006 Humana Press Inc. All rights of any nature whatsoever reserved.
Authors & Co-Authors
Rekhviashvili, Natela
South Africa, Johannesburg
University of the Witwatersrand
Stevens, Gwynneth
United States, New York
International Aids Vaccine Initiative
Scott, Lesley Erica
South Africa, Johannesburg
University of the Witwatersrand
Stevens, Wendy Susan
South Africa, Johannesburg
University of the Witwatersrand
Statistics
Citations: 11
Authors: 4
Affiliations: 2
Identifiers
Doi:
10.1385/MB:32:2:101
ISSN:
10736085
Research Areas
Infectious Diseases
Study Design
Cohort Study