A new strategy for studying in vitro the drug susceptibility of clinical isolates of human hepatitis B virus

Hepatology : Official Journal of the American Association for the Study of Liver Diseases
David DurantelFabien Zoulim

Abstract

Resistance of hepatitis B virus (HBV) to antivirals has become a major clinical problem. Our objective was to develop a new method for the cloning of naturally occurring HBV genomes and a phenotypic assay capable of assessing HBV drug susceptibility and DNA synthesis capacity in vitro. Viral DNA was extracted from sera and was amplified by polymerase chain reaction, and amplicons were cloned into vectors that enable, after cell transfection, the initiation of the intracellular HBV replication cycle. Single or multiple clones were used to transfect Huh7 cells. The viral DNA synthesis capacity and drug susceptibility were determined by measuring the level of intracellular DNA intermediate, synthesized in absence or presence of antiviral, using Southern blot analysis. We have developed, calibrated, then used this phenotypic assay to determine the drug susceptibility of HBV quasispecies isolated throughout the course of therapy from patients selected according to their mutation profile. A multiclonal and longitudinal analysis enabled us to measure the variation of drug susceptibility of different viral quasispecies by comparison of IC(50)/IC(90)s with standards. The presence of famciclovir- or lamivudine-induced mutations in the vi...Continue Reading

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Citations

Aug 18, 2012·Seminars in Immunopathology·Maura DandriJörg Petersen
Aug 10, 2007·Journal of Clinical Microbiology·Yuao ZhuKatyna Borroto-Esoda
Nov 17, 2012·World Journal of Gastroenterology : WJG·Maria HomsFrancisco Rodriguez-Frias
Feb 18, 2010·HIV Therapy·David M Iser, Sharon R Lewin
Nov 29, 2007·La Presse médicale·Fabien Zoulim
Aug 30, 2007·Journal of Acquired Immune Deficiency Syndromes : JAIDS·Yves Benhamou
Mar 10, 2015·International Journal of Molecular Sciences·Xiaosong LiAilong Huang
Jul 3, 2007·Expert Opinion on Emerging Drugs·Fabien Zoulim
Feb 4, 2012·Journal of Hepatology·Fabien Zoulim, Stephen Locarnini
Sep 10, 2009·Gastroenterology·Fabien Zoulim, Stephen Locarnini
Sep 9, 2008·Gastroentérologie Clinique Et Biologique·T AsselahP Marcellin
May 13, 2008·Gastroentérologie Clinique Et Biologique·F Bailly, F Zoulim
Nov 6, 2007·Clinics in Liver Disease·Claire Fournier, Fabien Zoulim
May 26, 2005·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Marie-Noëlle BrunelleFabien Zoulim
Jun 29, 2007·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Anna S LokUNKNOWN Hepatitis B Virus Drug Resistance Working Group
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Nov 21, 2007·Journal of Viral Hepatitis·F ZoulimA S Lok
Jan 8, 2008·Alimentary Pharmacology & Therapeutics·W F LeemansR A de Man
Nov 30, 2006·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Fabien Zoulim
Nov 5, 2011·Journal of Medical Virology·Jie-li HuAi-long Huang
Jan 18, 2005·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·Suzan D PasHubert G M Niesters
May 24, 2006·Antiviral Research·Fabien Zoulim
Feb 4, 2006·Journal of Hepatology·Tim ShawStephen Locarnini
Feb 8, 2006·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·David DurantelFabien Zoulim
Dec 14, 2005·Journal of Hepatology·Angelos HatzakisCatherine Haida
Dec 15, 2005·Journal of Hepatology·Fabien Zoulim
Feb 12, 2009·Liver International : Official Journal of the International Association for the Study of the Liver·Fabien ZoulimPaul Deny
Apr 17, 2016·Journal of Hepatology·Lena Allweiss, Maura Dandri
Apr 18, 2015·Clinical Microbiology and Infection : the Official Publication of the European Society of Clinical Microbiology and Infectious Diseases·R K MondalS Datta
Mar 9, 2012·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·Dong JiDongping Xu
Mar 12, 2008·Journal of Hepatology·Stéphanie VilletFabien Zoulim

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