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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

dc.contributor.authorDrosten, Christian
dc.contributor.authorPanning, Marcus
dc.contributor.authorDrexler, Jan Felixen_ZA
dc.contributor.authorHänsel, Florian
dc.contributor.authorPedroso, Celia
dc.contributor.authorYeats, Jane
dc.contributor.authorSamuel, Matthew
dc.contributor.authorLiedigk, Britta
dc.contributor.authorLippert, Ute
dc.contributor.authorStürmer, Martin
dc.contributor.authorDoerr, Hans Wilhelm
dc.contributor.authorBrites, Carlos
dc.contributor.authorPreiser, Wolfgang
dc.contributor.authorDe Souza Luna, Luciano Kleber
dc.date.accessioned2010-11-03T06:17:57Z
dc.date.available2010-11-03T06:17:57Z
dc.date.issued2006-04
dc.identifier.citationDrosten, C, Panning, M, Drexler, JF, Hansel, F, Pedroso, C, Yeats, J, De Souza Luna, LK, Samuel, M, Liedigk, B, Lippert, U, Sturmer, M, Doerr, HW, Brites, C & Preiser, W 2006, '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, vol. 52, pp. 1258-1266, DOI: 10.1373/clinchem.2006.066498en_ZA
dc.identifier.issn0009-9147 (print)
dc.identifier.issn1530-8561 (online)
dc.identifier.other10.1373/clinchem.2006.066498
dc.identifier.urihttp://hdl.handle.net/10019.1/4857
dc.descriptionBibliography
dc.description.abstractBACKGROUND: 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 log(10) compared with Amplicor in the 10(3)-10(4) 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.en_ZA
dc.format.extent8 p. : ill.
dc.language.isoen_ZAen_ZA
dc.publisherAmerican Association for Clinical Chemistryen_ZA
dc.subjectBrazilen_ZA
dc.subjectGenotypeen_ZA
dc.subjectGermanyen_ZA
dc.subjectHIV infections drug therapyen_ZA
dc.subjectHIV infections virologyen_ZA
dc.subjectHIV Long Terminal Repeaten_ZA
dc.subjectHIV-1* geneticsen_ZA
dc.subjectIndiaen_ZA
dc.subjectRNAen_ZA
dc.subjectViral analysisen_ZA
dc.subjectRegression analysisen_ZA
dc.subjectReverse Transcriptase Polymerase Chain Reaction economicsen_ZA
dc.subjectSouth Africaen_ZA
dc.subjectViral Loaden_ZA
dc.subjectRNAen_ZA
dc.subject.lcshHighly active retroviral therapy -- South Africa -- Testingen_ZA
dc.subject.lcshViral diseases therapy -- Testing -- Economic aspects -- Indiaen_ZA
dc.subject.lcshViral diseases therapy -- Testing -- Economic aspects -- Brazilen_ZA
dc.subject.lcshViral diseases therapy -- Testing -- Economic aspects -- Germanyen_ZA
dc.titleUltrasensitive 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 domainen_ZA
dc.typeArticleen_ZA
dc.rights.holderAmerican Association for Clinical Chemistryen_ZA


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