dc.contributor.author
Hauser, Andrea
dc.contributor.author
Kuecherer, Claudia
dc.contributor.author
Kunz, Andrea
dc.contributor.author
Dabrowski, Piotr Wojtek
dc.contributor.author
Radonić, Aleksandar
dc.contributor.author
Nitsche, Andreas
dc.contributor.author
Theuring, Stefanie
dc.contributor.author
Bannert, Norbert
dc.contributor.author
Sewangi, Julius
dc.contributor.author
Mbezi, Paulina
dc.contributor.author
Dugange, Festo
dc.contributor.author
Harms, Gundel
dc.contributor.author
Meixenberger, Karolin
dc.date.accessioned
2018-06-08T03:55:03Z
dc.date.available
2015-11-20T11:55:53.113Z
dc.identifier.uri
https://refubium.fu-berlin.de/handle/fub188/16202
dc.identifier.uri
http://dx.doi.org/10.17169/refubium-20386
dc.description.abstract
Background Pregnant HIV-infected women were screened for the development of
HIV-1 drug resistance after implementation of a triple-antiretroviral
transmission prophylaxis as recommended by the WHO in 2006. The study offered
the opportunity to compare amplicon-based 454 ultra-deep sequencing (UDS) and
allele-specific real-time PCR (ASPCR) for the detection of drug-resistant
minor variants in the HIV-1 reverse transcriptase (RT). Methods Plasma samples
from 34 Tanzanian women were previously analysed by ASPCR for key resistance
mutations in the viral RT selected by AZT, 3TC, and NVP (K70R, K103N, Y181C,
M184V, T215Y/F). In this study, the RT region of the same samples was
investigated by amplicon-based UDS for resistance mutations using the 454 GS
FLX System. Results Drug-resistant HIV-variants were identified in 69% (20/29)
of women by UDS and in 45% (13/29) by ASPCR. The absolute number of resistance
mutations identified by UDS was twice that identified by ASPCR (45 vs 24). By
UDS 14 of 24 ASPCR-detected resistance mutations were identified at the same
position. The overall concordance between UDS and ASPCR was 61.0% (25/41). The
proportions of variants quantified by UDS were approximately 2–3 times lower
than by ASPCR. Amplicon generation from samples with viral loads below 20,000
copies/ml failed more frequently by UDS compared to ASPCR (limit of detection
= 650 copies/ml), resulting in missing or insufficient sequence coverage.
Conclusions Both methods can provide useful information about drug-resistant
minor HIV-1 variants. ASPCR has a higher sensitivity than UDS, but is
restricted to single resistance mutations. In contrast, UDS is limited by its
requirement for high viral loads to achieve sufficient sequence coverage, but
the sequence information reveals the complete resistance patterns within the
genomic region analysed. Improvements to the UDS limit of detection are in
progress, and UDS could then facilitate monitoring of drug-resistant minor
variants in the HIV-1 quasispecies.
en
dc.rights.uri
http://creativecommons.org/licenses/by/4.0/
dc.subject.ddc
600 Technik, Medizin, angewandte Wissenschaften::610 Medizin und Gesundheit
dc.title
Comparison of 454 Ultra-Deep Sequencing and Allele-Specific Real-Time PCR with
Regard to the Detection of Emerging Drug-Resistant Minor HIV-1 Variants after
Antiretroviral Prophylaxis for Vertical Transmission
dc.type
Wissenschaftlicher Artikel
dcterms.bibliographicCitation
PLoS ONE. - 10 (2015), 10, Artikel Nr. e0140809
dcterms.bibliographicCitation.doi
10.1371/journal.pone.0140809
dcterms.bibliographicCitation.url
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0140809
refubium.affiliation
Charité - Universitätsmedizin Berlin
de
refubium.mycore.fudocsId
FUDOCS_document_000000023495
refubium.note.author
Der Artikel wurde in einer Open-Access-Zeitschrift publiziert.
refubium.resourceType.isindependentpub
no
refubium.mycore.derivateId
FUDOCS_derivate_000000005683
dcterms.accessRights.openaire
open access