Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
SHR Neuro Krebs Kardio Lipid Stoffw Microb
Stelzl, E; Troppan, KT; Winkler, M; Korn, K; Kessler, HH.
Optimized protocol for detection of HIV-1 drug mutations in patients with low viral load.
J VIROL METH. 2010; 168(1-2): 152-154.
Doi: 10.1016/j.jviromet.2010.05.007
Web of Science
PubMed
FullText
FullText_MUG
- Führende Autor*innen der Med Uni Graz
- Stelzl Evelyn
- Co-Autor*innen der Med Uni Graz
- Kessler Harald
- Prochazka Katharina
- Winkler Michaela
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- Abstract:
- The TRUGENE HIV-1 Genotyping Kit for HIV-1 drug resistance testing is limited by the need for samples with HIV-1 RNA loads of at least 1000 copies/mL. In order to enable sequencing of clinical samples with viral loads under 1000 copies/mL, an optimized automated sample preparation protocol on the VERSANT kPCR Sample Preparation (SP) Module was evaluated. In order to prove the concept of successful sequencing of low-titer clinical samples with the optimized protocol, a dilution series of a routine clinical sample was analyzed. Furthermore, 57 routine clinical samples with viral loads below 1000 HIV-1 RNA copies/mL were tested. Finally, samples obtained from two patients with low viral loads were tested retrospectively for HIV-1 drug resistances and results were compared with those of the preceding and the subsequent sample. The dilution containing 92 HIV-1 RNA copies/mL was the lowest yielding an analyzable sequence with the optimized protocol. When routine clinical samples with viral loads between 100 and 1000 HIV-1 RNA copies/mL were tested, a sequence was obtained in 90.5%. Samples with low viral load of two patients that could not be analyzed with the routine protocol showed identical drug mutations in both, the low viral-load and the subsequent samples. Together with the optimized automated sample preparation protocol, the TRUGENE HIV-1 Genotyping Kit allows successful sequencing of the majority of samples with HIV-1 RNA loads between 100 and 1000 copies/mL. Detection of resistance mutations in low viral-load samples may lead to an earlier optimized antiretroviral therapy.
- Find related publications in this database (using NLM MeSH Indexing)
- Anti-HIV Agents - pharmacology
- Automation - methods
- Drug Resistance, Viral -
- Genotype -
- HIV Infections - virology
- HIV-1 - drug effects
- Humans -
- Mutation, Missense -
- RNA, Viral - genetics
- Sensitivity and Specificity -
- Viral Load -
- Virology - methods
- Find related publications in this database (Keywords)
- Human immunodeficiency virus
- Drug resistance testing
- Viral load
- Sample preparation