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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
biochemistry, genetics and molecular biology
Ultrasensitive monitoring of HIV-1 viral load by a low-cost real-time reverse transcription-PCR assay with internal control for the 5′ long terminal repeat domain
Clinical Chemistry, Volume 52, No. 7, Year 2006
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Description
Background: Current HIV-1 viral-load assays are too expensive for resource-limited settings. In some countries, monitoring of antiretroviral therapy is now more expensive than treatment itself. In addition, some commercial assays have shown shortcomings in quantifying rare genotypes. Methods: We evaluated real-time reverse transcription-PCR with internal control targeting the conserved long terminal repeat (LTR) domain of HIV-1 on reference panels and patient samples from Brazil (n = 1186), South Africa (n = 130), India (n = 44), and Germany (n = 127). Results: The detection limit was 31.9 IU of HIV-1 RNA/mL of plasma (>95% probability of detection, Probit analysis). The internal control showed inhibition in 3.7% of samples (95% confidence interval, 2.32%-5.9%; n = 454; 40 different runs). Comparative qualitative testing yielded the following: Roche Amplicor vs LTR assay (n = 431 samples), 51.7% vs 65% positives; Amplicor Ultrasensitive vs LTR (n = 133), 81.2% vs 82.7%; BioMerieux NucliSens HIV-1 QT (n = 453), 60.5% vs 65.1%; Bayer Versant 3.0 (n = 433), 57.7% vs 55.4%; total (n = 1450), 59.0% vs 63.8% positives. Intra-/interassay variability at medium and near-negative concentrations was 18%-51%. The quantification range was 50-10 000 000 IU/mL. Viral loads for subtypes A-D, F-J, AE, and AG yielded mean differences of 0.31 log10 compared with Amplicor in the 103-104 IU/mL range. HIV-1 N and O were not detected by Amplicor, but yielded up to 180 180.00 IU/mL in the LTR assay. Viral loads in stored samples from all countries, compared with Amplicor, NucliSens, or Versant, yielded regression line slopes (SD) of 0.9 (0.13) (P <0.001 for all). Conclusions: This method offers all features of commercial assays and covers all relevant genotypes. It could allow general monitoring of antiretroviral therapy in resource-limited settings. © 2006 American Association for Clinical Chemistry.
Available Materials
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https://efashare.b-cdn.net/share/pmc/articles/PMC7108179/bin/clinchem.2006.066498-2.doc
Authors & Co-Authors
Drosten, C.
Germany, Hamburg
Bernhard Nocht Institut Fur Tropenmedizin Hamburg
Panning, Marcus
Germany, Hamburg
Bernhard Nocht Institut Fur Tropenmedizin Hamburg
Drexler, Jan Felix
Germany, Hamburg
Bernhard Nocht Institut Fur Tropenmedizin Hamburg
Brazil, Salvador
Universidade Federal da Bahia
Hänsel, Florian
Germany, Hamburg
Bernhard Nocht Institut Fur Tropenmedizin Hamburg
Pedroso, Célia
Brazil, Salvador
Universidade Federal da Bahia
Yeats, Jane
South Africa, Cape Town
University of Cape Town
De Souza Luna, Luciano Kleber
Germany, Hamburg
Bernhard Nocht Institut Fur Tropenmedizin Hamburg
Samuel, Matthew
India, Chennai
Tamil Nadu Medical University
Liedigk, Britta
Germany, Hamburg
Bernhard Nocht Institut Fur Tropenmedizin Hamburg
Lippert, Ute
Germany, Hamburg
Bernhard Nocht Institut Fur Tropenmedizin Hamburg
Stürmer, Martin
Germany, Frankfurt am Main
Goethe-universität Frankfurt am Main
Doérr, Hans Wilhelm
Germany, Frankfurt am Main
Goethe-universität Frankfurt am Main
Brites, Carlos R.
Brazil, Salvador
Universidade Federal da Bahia
Preiser, Wolfgang
Germany, Frankfurt am Main
Goethe-universität Frankfurt am Main
South Africa, Stellenbosch
Stellenbosch University
Statistics
Citations: 116
Authors: 14
Affiliations: 6
Identifiers
Doi:
10.1373/clinchem.2006.066498
ISSN:
00099147
Research Areas
Health System And Policy
Infectious Diseases
Study Approach
Qualitative
Study Locations
South Africa