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Journal of Clinical Microbiology, April 2001, p. 1456-1459, Vol. 39, No. 4
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.4.1456-1459.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Monitoring the Emergence of Hepatitis B Virus Polymerase Gene Variants during Lamivudine Therapy Using the LightCycler

Simon A. Whalley,1,2 Dave Brown,1 Chong Gee Teo,2 Geoffrey M. Dusheiko,1 and Nicholas A. Saunders2,*

Department of Medicine, Royal Free and University College Medical School,1 and Hepatitis and Retrovirus Laboratory, Central Public Health Laboratory,2 London, United Kingdom

Received 6 June 2000/Returned for modification 21 September 2000/Accepted 26 January 2001

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.

* Corresponding author. Mailing address: Sexually Transmitted and Blood Borne Virus Laboratory, PHLS Central Public Health Laboratory, 61 Colindale Ave., London NW9 5HT, United Kingdom. Phone: 020 8200 4400. Fax: 020 8200 1569. E-mail: nsaunders{at}phls.org.uk.

Journal of Clinical Microbiology, April 2001, p. 1456-1459, Vol. 39, No. 4
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.4.1456-1459.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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