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Monitoring the emergence of hepatitis B virus polymerase gene variants during lamivudine therapy using the LightCycler

Whalley, SA; Brown, D; Teo, CG; Dusheiko, GM; Saunders, NA; (2001) Monitoring the emergence of hepatitis B virus polymerase gene variants during lamivudine therapy using the LightCycler. Journal of Clinical Microbiology , 39 (4) 1456 - 1459.

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Abstract

Treatment of chronic hepatitis B virus (HBV) infection with lamivudine is associated with the appearance in the circulation of HBV variants with mutations in the YMDD (tyrosine, methionine, aspartate, aspartate) motif of the polymerase gene. Fluorometric real-time PCR with the LightCycler assay was used for the detection of resistant variants. Differences in the hybridization melting curve kinetics of probes bound to the sequences encoding the wild-type or the mutant YMDD motifs (YIDD or YVDD in which the methionine residue is altered to an isoleucine or a valine, respectively) distinguished the single-base changes responsible for the resistance phenotype. The LightCycler probe hybridization assay was applied to 40 serum specimens from 19 patients, and the results were correlated with the nucleotide sequences determined for the corresponding PCR products. All three variants could be identified in the specimens. PCR clones obtained from four patients early in the course and prior to lamivudine therapy were investigated for the appearance of YIDD and YVDD variants with the LightCycler assay. In one patient, a transient appearance of the YIDD variant was observed 6 weeks into therapy. Subsequently, after 11 months of lamivudine therapy, the YVDD variant emerged in that patient

Type:Article
Title:Monitoring the emergence of hepatitis B virus polymerase gene variants during lamivudine therapy using the LightCycler
Additional information:UI - 21179269 LA - eng RN - 0 (Antiviral Agents) RN - 0 (Gene Products, pol) RN - 0 (P protein, hepatitis B virus) RN - 134678-17-4 (Lamivudine) PT - Journal Article DA - 20010403 IS - 0095-1137 SB - IM CY - United States
Keywords:appearance, Applied, ASSAY, Blood, Chronic, CHRONIC HEPATITIS, chronic hepatitis B, CIRCULATION, CLONE, CLONES, detection, difference, English, GENE, HBV, Hepatitis, hepatitis B, Hepatitis B Virus, HIGH-LEVEL RESISTANCE, HYBRIDIZATION, IDENTIFICATION, Infection, Isoleucine, kinetic, Kinetics, Lamivudine, LAMIVUDINE THERAPY, LIVER- TRANSPLANTATION, Methionine, microbiology, monitoring, MOTIFS, Mutant, Mutation, MUTATIONS, N, Nucleotide, Nucleotide Sequence, NUCLEOTIDE-SEQUENCE, Patient, patients, PCR, Phenotype, polymerase, PROBE, PROBES, product, PRODUCTS, real-time, Resistance, Result, retrovirus, REVERSE-TRANSCRIPTASE, selection, SEQUENCE, SEQUENCES, SERA, serum, SINGLE, SPECIMENS, ST, THERAPIES, therapy, treatment, TRIAL, TYPE-1, Tyrosine, USA, Valine, VARIANT, VARIANTS, VIRUS, WILD-TYPE, AGENT, AGENTS, Antiviral Agents, appearance, Drug Resistance, Microbial, drug therapy, enzymology, Fluorescence, GENE PRODUCT, Gene Products, pol, GENE-PRODUCT, genetics, IM, LA, Methods, pharmacology, Polymerase Chain Reaction, PROTEIN, STATE, STATES, Support, Non-U.S.Gov't, therapeutic use, united, United States, UNITED-STATES, virology

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