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Ultra-deep pyrosequencing of hepatitis B virus quasispecies from nucleoside and nucleotide reverse-transcriptase inhibitor (NRTI)-treated patients and NRTI-naive patients
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2009 (English)In: Journal of Infectious Diseases, ISSN 0022-1899, Vol. 199, no 9, 1275-1285 p.Article in journal (Refereed) Published
Abstract [en]

The dynamics of emerging nucleoside and nucleotide reverse-transcriptase inhibitor (NRTI) resistance in hepatitis B virus (HBV) are not well understood because standard dideoxynucleotide direct polymerase chain reaction (PCR) sequencing assays detect drug-resistance mutations only after they have become dominant. To obtain insight into NRTI resistance, we used a new sequencing technology to characterize the spectrum of low-prevalence NRTI-resistance mutations in HBV obtained from 20 plasma samples from 11 NRTI-treated patients and 17 plasma samples from 17 NRTI-naive patients, by using standard direct PCR sequencing and ultra-deep pyrosequencing (UDPS). UDPS detected drug-resistance mutations that were not detected by PCR in 10 samples from 5 NRTI-treated patients, including the lamivudine-resistance mutation V173L (in 5 samples), the entecavir-resistance mutations T184S (in 2 samples) and S202G (in 1 sample), the adefovir-resistance mutation N236T (in 1 sample), and the lamivudine and adefovir-resistance mutations V173L, L180M, A181T, and M204V (in 1 sample). G-to-A hypermutation mediated by the apolipoprotein B mRNA editing enzyme, catalytic polypeptide-like family of cytidine deaminases was estimated to be present in 0.6% of reverse-transcriptase genes. Genotype A coinfection was detected by UDPS in each of 3 patients in whom genotype G virus was detected by direct PCR sequencing. UDPS detected low-prevalence HBV variants with NRTI-resistance mutations, G-to-A hypermutation, and low-level dual genotype infection with a sensitivity not previously possible.

Place, publisher, year, edition, pages
University of Chicago Press, 2009. Vol. 199, no 9, 1275-1285 p.
National Category
Biological Sciences
URN: urn:nbn:se:kth:diva-137244DOI: 10.1086/597808ISI: 000265035500007OAI: diva2:678512
15th Conference on Retroviruses and Opportunistic Infections,Boston, MA,FEB 03-06, 2008

QC 20140924

Available from: 2013-12-12 Created: 2013-12-12 Last updated: 2014-09-24Bibliographically approved

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Babrzadeh, Farbod
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