1887

Abstract

Monitoring hepatitis B virus (HBV) mutants periodically during nucleoside analogue treatment is of great clinical significance, particularly in persistently HBV DNA-positive patients. However, few studies have investigated the dynamic changes of HBV YMDD (Tyr–Met–Asp–Asp) and YVDD (Tyr–Val–Asp–Asp) populations in chronic hepatitis B (CHB) patients whilst undergoing lamivudine (LMV) treatment. In this study, we sought to investigate the dynamic changes of HBV YMDD and YVDD variants by ultrasensitive real-time amplification refractory mutation system quantitative PCR (RT-ARMS-qPCR) and evaluate its significance for changes in the treatment of CHB patients. RT-ARMS-qPCR was established and evaluated with standard recombinant plasmids. Fifteen CHB patients receiving LMV (100 mg daily) were consecutively recruited and followed up for 60 weeks. Serum samples were obtained from each patient at baseline and every 12 weeks. The total HBV DNA, HBV YMDD DNA and YVDD DNA levels were measured using RT-ARMS-qPCR at all given time points after treatment. Routine liver biochemistry parameters, including aspartate aminotransferase and alanine aminotransferase, were also measured every 12 weeks. The linear range of the assay was between 1×10 and 1×10 copies ml. The low detection limit was 1×10 copies ml. After 60 weeks of LMV treatment, nine patients experienced virological breakthrough. The YVDD variant could be detected 12–48 weeks before virological breakthrough. The YVDD variant was detected as the predominant population (range 69.4–100 %) in patients by the time virological breakthrough appeared. We concluded that RT-ARMS-qPCR was sensitive for the detection and quantification of low levels of HBV mutation. Periodic detection of HBV YM(V)DD every 12 weeks during LMV treatment is helpful for therapeutic decision making.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.000022
2015-03-01
2019-11-18
Loading full text...

Full text loading...

/deliver/fulltext/jmm/64/3/237.html?itemId=/content/journal/jmm/10.1099/jmm.0.000022&mimeType=html&fmt=ahah

References

  1. Akarsu M. , Sengonul A. , Tankurt E. , Sayiner A. A. , Topalak O. , Akpinar H. , Abacioglu Y. H. . ( 2006; ). YMDD motif variants in inactive hepatitis B carriers detected by Inno-Lipa HBV DR assay. . J Gastroenterol Hepatol 21:, 1783–1788. [CrossRef] [PubMed]
    [Google Scholar]
  2. Akuta N. , Suzuki F. , Kobayashi M. , Tsubota A. , Suzuki Y. , Hosaka T. , Someya T. , Kobayashi M. , Saitoh S. et al. ( 2003; ). The influence of hepatitis B virus genotype on the development of lamivudine resistance during long-term treatment. . J Hepatol 38:, 315–321. [CrossRef] [PubMed]
    [Google Scholar]
  3. Buti M. , Cotrina M. , Valdes A. , Jardi R. , Rodriguez-Frias F. , Esteban R. . ( 2002; ). Is hepatitis B virus subtype testing useful in predicting virological response and resistance to lamivudine?. J Hepatol 36:, 445–446. [CrossRef] [PubMed]
    [Google Scholar]
  4. Chang T. T. , Lai C. L. , Kew Yoon S. , Lee S. S. , Coelho H. S. , Carrilho F. J. , Poordad F. , Halota W. , Horsmans Y. et al. ( 2010; ). Entecavir treatment for up to 5 years in patients with hepatitis B e antigen-positive chronic hepatitis B. . Hepatology 51:, 422–430. [CrossRef] [PubMed]
    [Google Scholar]
  5. Chen L. , Zhang Q. , Yu D. M. , Wan M. B. , Zhang X. X. . ( 2009; ). Early changes of hepatitis B virus quasispecies during lamivudine treatment and the correlation with antiviral efficacy. . J Hepatol 50:, 895–905. [CrossRef] [PubMed]
    [Google Scholar]
  6. European Association for the Study of the Liver ( 2012; ). EASL Clinical Practice Guidelines: Management of chronic hepatitis B virus infection. . J Hepatol 57:, 167–185.[PubMed] [CrossRef]
    [Google Scholar]
  7. Feng B. , Wei L. , Chen M. , Wang L. . ( 2008; ). Dynamic changes of hepatitis B virus polymerase gene including YMDD motif in lamivudine-treated patients with chronic hepatitis B. . Microbiol Res 163:, 487–492. [CrossRef] [PubMed]
    [Google Scholar]
  8. Hsieh T. H. , Tseng T. C. , Liu C. J. , Lai M. Y. , Chen P. J. , Hsieh H. L. , Chen D. S. , Kao J. H. . ( 2009; ). Hepatitis B virus genotype B has an earlier emergence of lamivudine resistance than genotype C. . Antivir Ther 14:, 1157–1163. [CrossRef] [PubMed]
    [Google Scholar]
  9. Keeffe E. B. , Dieterich D. T. , Pawlotsky J. M. , Benhamou Y. . ( 2008; ). Chronic hepatitis B: preventing, detecting, and managing viral resistance. . Clin Gastroenterol Hepatol 6:, 268–274. [CrossRef] [PubMed]
    [Google Scholar]
  10. Kobayashi S. , Ide T. , Sata M. . ( 2001; ). Detection of YMDD motif mutations in some lamivudine-untreated asymptomatic hepatitis B virus carriers. . J Hepatol 34:, 584–586. [CrossRef] [PubMed]
    [Google Scholar]
  11. Li Z. G. , Chen L. Y. , Huang J. , Qiao P. , Qiu J. M. , Wang S. Q. . ( 2005; ). Quantification of the relative levels of wild-type and lamivudine-resistant mutant virus in serum of HBV-infected patients using microarray. . J Viral Hepat 12:, 168–175. [CrossRef] [PubMed]
    [Google Scholar]
  12. Liu Y. , Xu Z. , Wang Y. , Li X. , Liu L. , Chen L. , Xin S. , Xu D. . ( 2014; ). rtM204Q may serve as a novel lamivudine-resistance-associated mutation of hepatitis B virus. . PLoS One 9:, e89015. [CrossRef] [PubMed]
    [Google Scholar]
  13. Lok A. S. , McMahon B. J. . ( 2007; ). Chronic hepatitis B. . Hepatology 45:, 507–539. [CrossRef] [PubMed]
    [Google Scholar]
  14. Pallier C. , Castéra L. , Soulier A. , Hézode C. , Nordmann P. , Dhumeaux D. , Pawlotsky J. M. . ( 2006; ). Dynamics of hepatitis B virus resistance to lamivudine. . J Virol 80:, 643–653. [CrossRef] [PubMed]
    [Google Scholar]
  15. Wu F. , Wu M. J. , Zhuge X. L. , Zhu S. M. , Zhu B. . ( 2012; ). Correlation of the occurrence of YMDD mutations with HBV genotypes, HBV-DNA levels, and HBeAg status in Chinese patients with chronic hepatitis B during lamivudine treatment. . Hepatobiliary Pancreat Dis Int 11:, 172–176. [CrossRef] [PubMed]
    [Google Scholar]
  16. Zeng Y. , Li D. , Wang W. , Su M. , Lin J. , Chen H. , Jiang L. , Chen J. , Yang B. , Ou Q. . ( 2014; ). Establishment of real time allele specific locked nucleic acid quantitative PCR for detection of HBV YIDD (ATT) mutation and evaluation of its application. . PLoS One 9:, e90029. [CrossRef] [PubMed]
    [Google Scholar]
  17. Zhao J. , Guo Y. , Yan Z. , Liang P. , Zhang J. , Liu Y. . ( 2012; ). The natural YMDD mutations of hepatitis B virus in Western China. . Scand J Infect Dis 44:, 44–47. [CrossRef] [PubMed]
    [Google Scholar]
  18. Zoulim F. , Locarnini S. . ( 2009; ). Hepatitis B virus resistance to nucleos(t)ide analogues. . Gastroenterology 137:, 1593–1608, e1–e2. [CrossRef] [PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.000022
Loading
/content/journal/jmm/10.1099/jmm.0.000022
Loading

Data & Media loading...

Supplements

Supplementary Data



PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error