PMID- 19559728
OWN - NLM
STAT- MEDLINE
DCOM- 20091109
LR  - 20131121
IS  - 1879-0984 (Electronic)
IS  - 0166-0934 (Linking)
VI  - 161
IP  - 2
DP  - 2009 Nov
TI  - Detection and identification of U69 gene mutations encoded by 
      ganciclovir-resistant human herpesvirus 6 using denaturing high-performance 
      liquid chromatography.
PG  - 223-30
LID - 10.1016/j.jviromet.2009.06.016 [doi]
AB  - A denaturing high-performance liquid chromatography (dHPLC) assay was developed 
      to detect antiviral drug-resistance mutations of human herpesvirus 6 (HHV-6). 
      Recombinant baculoviruses were created that contained wild-type and mutant forms 
      of the HHV-6 U69 gene, which determines sensitivity to the antiviral drug 
      ganciclovir (GCV). The mutations causing GCV resistance in HHV-6 U69 were 
      single-base mutations adapted from known GCV-resistant DNA sequences of HCMV, and 
      their ability to confer GCV resistance on recombinant baculoviruses was 
      confirmed. Six characterized mutant sequences, including one reported previously 
      that encodes a GCV-sensitive kinase-activity mutant, were used. DNA was 
      extracted, and the levels of homoduplex and heteroduplex DNA in the PCR products 
      from mixed wild-type and mutant viral DNAs were determined using dHPLC. The 
      optimized assay could distinguish the different mutants, and could detect mutants 
      representing only 10% of the DNAs. The new assay with dHPLC readout permitted the 
      rapid (4 h), objective, and reproducible detection of HHV-6 drug-resistance 
      mutations.
FAU - Nakano, Kazushi
AU  - Nakano K
AD  - Department of Infectious Disease Control, G-5, Graduate School of Medicine, Osaka 
      University, Suita, Osaka, Japan.
FAU - Nishinaka, Kazuko
AU  - Nishinaka K
FAU - Tanaka, Tatsuya
AU  - Tanaka T
FAU - Ohshima, Atsushi
AU  - Ohshima A
FAU - Sugimoto, Nakaba
AU  - Sugimoto N
FAU - Isegawa, Yuji
AU  - Isegawa Y
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20090625
PL  - Netherlands
TA  - J Virol Methods
JT  - Journal of virological methods
JID - 8005839
RN  - 0 (Antiviral Agents)
RN  - 0 (DNA, Viral)
RN  - P9G3CKZ4P5 (Ganciclovir)
SB  - IM
MH  - Antiviral Agents/*therapeutic use
MH  - Base Sequence
MH  - Chromatography, High Pressure Liquid/methods
MH  - DNA, Viral/chemistry/genetics
MH  - Drug Resistance, Viral/*genetics
MH  - Ganciclovir/*therapeutic use
MH  - *Genes, Viral
MH  - Herpesvirus 6, Human/chemistry/drug effects/*genetics
MH  - Humans
MH  - Molecular Sequence Data
MH  - *Point Mutation
MH  - Reproducibility of Results
MH  - Roseolovirus Infections/drug therapy/*virology
MH  - Sensitivity and Specificity
EDAT- 2009/06/30 09:00
MHDA- 2009/11/10 06:00
CRDT- 2009/06/30 09:00
PHST- 2009/02/24 00:00 [received]
PHST- 2009/06/10 00:00 [revised]
PHST- 2009/06/16 00:00 [accepted]
PHST- 2009/06/30 09:00 [entrez]
PHST- 2009/06/30 09:00 [pubmed]
PHST- 2009/11/10 06:00 [medline]
AID - S0166-0934(09)00301-2 [pii]
AID - 10.1016/j.jviromet.2009.06.016 [doi]
PST - ppublish
SO  - J Virol Methods. 2009 Nov;161(2):223-30. doi: 10.1016/j.jviromet.2009.06.016. 
      Epub 2009 Jun 25.