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.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.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.publisherAmerican Association for Clinical Chemistryen_ZA
dc.subjectHIV infections drug therapyen_ZA
dc.subjectHIV infections virologyen_ZA
dc.subjectHIV Long Terminal Repeaten_ZA
dc.subjectHIV-1* geneticsen_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.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.rights.holderAmerican Association for Clinical Chemistryen_ZA

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