Publikation

Detection of drug-resistant HIV minorities in clinical specimens and therapy failure

Wissenschaftlicher Artikel/Review - 01.03.2008

Bereiche
PubMed
DOI

Zitation
Louvel S, Battegay M, Vernazza P, Bregenzer T, Klimkait T, Hamy F, Swiss HIV Cohort Study. Detection of drug-resistant HIV minorities in clinical specimens and therapy failure. HIV medicine 2008; 9:133-41.
Art
Wissenschaftlicher Artikel/Review (Englisch)
Zeitschrift
HIV medicine 2008; 9
Veröffentlichungsdatum
01.03.2008
ISSN (Druck)
1464-2662
Seiten
133-41
Kurzbeschreibung/Zielsetzung

OBJECTIVE: Particularly for therapy-experienced patients, resistance assessment by genotypic or phenotypic methods produces discordances. This study seeks proof that differences may arise from the fact that genotyping produces a single summary sequence whereas replicative phenotyping (rPhenotyping) functionally detects and assigns resistances in mixed HIV populations. METHODS: For validation, defined mixes of wild-type and M184V mutant were analysed by rPhenotyping or standard genotyping. Allele-specific and quantitative polymerase chain reaction (PCR) set detection and quantification limits for minor virus populations in vitro and in authentic clinical samples showing geno-/pheno-discrepant lamivudine resistance. RESULTS: Allele-specific and real-time PCR methods detected down to 0.3% of mutant M184V. The functional assessment was sensitive enough to reveal <1% of mutant M184V in mixed samples. Also in discordant samples from the diagnostic routine, in which rPhenotyping had identified drug resistance, real-time PCR confirmed minute amounts of mutant M184V. CONCLUSION: By utilizing the replication dynamics of HIV under drug pressure, a rPhenotyping format potently reveals relevant therapy-resistant minority species, even of HIV known to possess reduced replicative fitness. With its rapid turnaround of 8 days and its high sensitivity, our rPhenotyping system may be a valuable diagnostic tool for detecting the early emergence of therapy-threatening HIV minorities or the persistence of residual resistant virus.