PMID- 21056552
OWN - NLM
STAT- MEDLINE
DCOM- 20110324
LR  - 20220408
IS  - 1873-3492 (Electronic)
IS  - 0009-8981 (Linking)
VI  - 412
IP  - 3-4
DP  - 2011 Jan 30
TI  - Variable influence of mutational patterns in reverse-transcriptase domain on 
      replication capacity of hepatitis B virus isolates from antiviral-experienced 
      patients.
PG  - 305-13
LID - 10.1016/j.cca.2010.10.028 [doi]
AB  - BACKGROUND: Various mutations in reverse-transcriptase domain (RT) of hepatitis B 
      virus (HBV) polymerase may develop during antiviral therapy. The influence of 
      these mutational patterns on HBV replication capacity remains to be fully 
      clarified. METHODS: Nine clones containing complete HBV genomes were isolated 
      from 5 patients with chronic hepatitis B who had received antiviral treatment. 
      Viral replication capacity was measured by quantitation of HBV replicative 
      intermediates using vector-free transfer of paired mutant and wild-type HBV 
      genomes into human hepatoma cell lines HepG2 and Huh7. HBV pgRNA was quantitated 
      by real-time PCR and Southern blot analysis. RESULTS: A real-time PCR assay with 
      high sensitivity and small variation was developed for quantitation of HBV 
      replicative intermediates. Compared to wild-type counterpart, mutant rtL217P 
      produced 1.98-fold higher replicative intermediate level, and mutant 
      rtM204I+rtL217P increased the replicative intermediate level to 1.20 fold. Other 
      mutational patterns (rtV173M, rtA181S/V, rtM204I, rtQ215H, rtL229M, rtN238H, 
      rtV84M+rtA181S+rtM204I, rtV84M+rtM204I, rtA181S+rtM204I, rtA181V+rtL229M, 
      rtQ215H+rtN238H) reduced viral replication capacity to different extents. 
      CONCLUSIONS: The study offers a practical measurement assay and novel information 
      for replication features of mutant strains; especially, rtL217P substitution 
      likely represents an energetic replication-compensatory mutation.
CI  - Copyright (c) 2010 Elsevier B.V. All rights reserved.
FAU - Ji, Dong
AU  - Ji D
AD  - Viral Hepatitis Research Laboratory, Institute of Infectious Diseases, Beijing 
      302 Hospital, Beijing, China.
FAU - Liu, Yan
AU  - Liu Y
FAU - Si, Lan-Lan
AU  - Si LL
FAU - Li, Le
AU  - Li L
FAU - Chen, Guo-Feng
AU  - Chen GF
FAU - Xin, Shao-Jie
AU  - Xin SJ
FAU - Zhao, Jin-Min
AU  - Zhao JM
FAU - Xu, Dongping
AU  - Xu D
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20101105
PL  - Netherlands
TA  - Clin Chim Acta
JT  - Clinica chimica acta; international journal of clinical chemistry
JID - 1302422
RN  - 0 (Antiviral Agents)
RN  - 0 (RNA, Viral)
RN  - EC 2.7.7.49 (RNA-Directed DNA Polymerase)
SB  - IM
MH  - Antiviral Agents/*therapeutic use
MH  - Calibration
MH  - Genome, Viral/genetics
MH  - Genotype
MH  - Hep G2 Cells
MH  - Hepatitis B virus/*enzymology/genetics/*physiology
MH  - Hepatitis B, Chronic/drug therapy/virology
MH  - Humans
MH  - Intracellular Space/metabolism
MH  - Mutagenesis, Site-Directed
MH  - *Mutation
MH  - Polymerase Chain Reaction
MH  - Protein Structure, Tertiary
MH  - RNA, Viral/metabolism
MH  - RNA-Directed DNA Polymerase/*chemistry/genetics/*metabolism
MH  - Reproducibility of Results
MH  - *Virus Replication
EDAT- 2010/11/09 06:00
MHDA- 2011/03/25 06:00
CRDT- 2010/11/09 06:00
PHST- 2010/02/08 00:00 [received]
PHST- 2010/10/10 00:00 [revised]
PHST- 2010/10/29 00:00 [accepted]
PHST- 2010/11/09 06:00 [entrez]
PHST- 2010/11/09 06:00 [pubmed]
PHST- 2011/03/25 06:00 [medline]
AID - S0009-8981(10)00680-7 [pii]
AID - 10.1016/j.cca.2010.10.028 [doi]
PST - ppublish
SO  - Clin Chim Acta. 2011 Jan 30;412(3-4):305-13. doi: 10.1016/j.cca.2010.10.028. Epub 
      2010 Nov 5.