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Journal of Clinical Microbiology, October 2002, p. 3729-3734, Vol. 40, No. 10
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.10.3729-3734.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Monitoring Drug Resistance in Chronic Hepatitis B Virus (HBV)-Infected Patients during Lamivudine Therapy: Evaluation of Performance of INNO-LiPA HBV DR Assay

Anna S. F. Lok,1 Fabien Zoulim,2 Stephen Locarnini,3 Alessandra Mangia,4 Grazia Niro,4 Hilde Decraemer,5 Geert Maertens,5 Frank Hulstaert,5 Karen De Vreese,5 and Erwin Sablon5*

Division of Gastroenterology, University of Michigan Medical Center, Ann Arbor, Michigan 48109,1 INSERM Unit 271, Institut Universitaire de France, 69003 Lyon, France,2 Research and Molecular Development, WHO Collaborating Centers for Virus Reference and Research and for Biosafety, The Victorian Infectious Diseases Reference Laboratory, North Melbourne 3051, Australia,3 Divisione di Gastroenterologia, Ospedale Generale Regionale "Casa Sollievo della Sofferenza," 71013 San Giovanni Rotondo (Fg), Italy,4 Innogenetics N.V., B-9052 Ghent, Belgium5

Received 15 February 2002/ Returned for modification 18 May 2002/ Accepted 20 July 2002

Sensitive and early detection of emerging hepatitis B virus (HBV) drug resistance may not only help monitor the viral dynamics associated with lamivudine treatment but could also improve therapeutic decision making. This is especially important when new antivirals effective against lamivudine-resistant HBV become available. A total of 159 serum samples from 33 chronic HBV patients receiving lamivudine treatment were analyzed at four centers for the presence of lamivudine-resistant mutations at codons 528 [180] (proposed revised nomenclature according to Stuyver et al. [Hepatology 33:751-757, 2001] shown in brackets), 552 [204], and 555 [207] of the HBV polymerase. Sequencing data were compared with results generated by the INNO-LiPA HBV DR line probe assay (LiPA), an assay based on reverse hybridization of amplified HBV DNA fragments with specific nucleotide probes immobilized on nitrocellulose strips. LiPA provided at least the same information as sequencing for 97.5% of all codons analyzed for codon 528 [180], 95% for codon 552 [204], and 100% for codon 555 [207]. The most common reason for discrepant or indeterminate results (0.4% and 1.5%, respectively) in a small percentage of the population tested could be attributed to polymorphisms not yet covered by LiPA probes. In at least five patients, a mutant could be detected earlier by LiPA than by sequencing. In 15 patients, LiPA detected mixed wild-type and mutant virus populations before viral breakthrough. These results demonstrate that INNO-LiPA HBV DR is a highly sensitive and easily applicable assay for the detection and monitoring of lamivudine-resistant mutations in chronic hepatitis B patients and that the assay is more sensitive than sequencing in detecting mixed mutant and wild-type sequences.

* Corresponding author. Mailing address: Innogenetics N.V., Industriepark Zwijnaarde 7/4, B-9052 Ghent, Belgium. Phone: 32 9 2410804. Fax: 32 9 2410907. E-mail: erwin_sablon{at}innogenetics.be.

Journal of Clinical Microbiology, October 2002, p. 3729-3734, Vol. 40, No. 10
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.10.3729-3734.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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