| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
1 Division of Virology, National Institute for Medical Research, Mill Hill, London NW7 1AA
and2 Department of Molecular Sciences, Wellcome Research Laboratories, Beckenham, Kent BR3 3BS, U.K.
We have applied the polymerase chain reaction (PCR) technique to analyse mutations in the thymidine kinase (TK) gene of varicella-zoster virus (VZV) associated with resistance to the 5-bromovinyl (BVaraU) and 5-propynyl (PYaraU) analogues of arabinofuranosyl deoxyuridine. The results from this study allow three clear conclusions to be drawn. Firstly, the technique clearly shows that populations of VZV derived from plaque purification were truly clonal only when the plaques were initiated from cell-free virus (representing a tiny fraction of infectious virus) and plaques initiated by infected cells contained a mixture of variants. Secondly, despite the background mutations caused by errors of the Taq DNA polymerase, mutations relevant to drug resistance can easily be distinguished. The BVaraU-resistant mutant, 7-1, contained an aspartic acid to asparagine mutation at residue 18 and a single base deletion (position 65298 of the VZV DNA sequence), resulting in a frameshift and premature termination of the polypeptide chain, was found in the BVaraU-resistant mutant YSR. PYaraU-resistant virus populations contained viruses with one or more of three independent mutations, i.e. single base substitutions resulting in mutations from leucine to proline at residue 92, histidine to arginine at residue 97 and a deletion of 20bp (residues 65135 to 65154). Finally, the technique has uncovered novel sites in the virus TK associated with drug resistance. We conclude that in vitro amplification using the PCR combined with cloning and sequencing is a relatively rapid method for identifying mutations in small virus populations even when they are not homogeneous.
Received 18 September 1990;
accepted 2 November 1990.
This article has been cited by other articles:
|
|
 |
|
  M. T. M. Vossen, M.-R. Gent, K. M. C. Peters, P. M. E. Wertheim-van Dillen, K. M. Dolman, A. van Breda, R. A. W. van Lier, and T. W. Kuijpers Persistent Detection of Varicella-Zoster Virus DNA in a Previously Healthy Child after Severe Chickenpox J. Clin. Microbiol., November 1, 2005; 43(11): 5614 - 5621. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Grose, S. Tyler, G. Peters, J. Hiebert, G. M. Stephens, W. T. Ruyechan, W. Jackson, J. Storlie, and G. A. Tipples Complete DNA Sequence Analyses of the First Two Varicella-Zoster Virus Glycoprotein E (D150N) Mutant Viruses Found in North America: Evolution of Genotypes with an Accelerated Cell Spread Phenotype J. Virol., July 1, 2004; 78(13): 6799 - 6807. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Suzutani, M. Saijo, M. Nagamine, M. Ogasawara, and M. Azuma Rapid Phenotypic Characterization Method for Herpes Simplex Virus and Varicella-Zoster Virus Thymidine Kinases To Screen for Acyclovir-Resistant Viral Infection J. Clin. Microbiol., May 1, 2000; 38(5): 1839 - 1844. [Abstract] [Full Text]
|
|
|
|
 |
|
 R. Sahli, G. Andrei, C. Estrade, R. Snoeck, and P. R. A. Meylan A Rapid Phenotypic Assay for Detection of Acyclovir-Resistant Varicella-Zoster Virus with Mutations in the Thymidine Kinase Open Reading Frame Antimicrob. Agents Chemother., April 1, 2000; 44(4): 873 - 878. [Abstract] [Full Text]
|
|
|
|
|
|
N. Ono, S. Iwayama, K. Suzuki, T. Sekiyama, H. Nakazawa, T. Tsuji, M. Okunishi, T. Daikoku, and Y. Nishiyama Mode of Action of (1'S,2'R)-9-{[1',2'-Bis(hydroxymethyl) cycloprop-1'-yl]methyl}guanine (A-5021) against Herpes Simplex Virus Type 1 and Type 2 and Varicella-Zoster Virus Antimicrob. Agents Chemother., August 1, 1998; 42(8): 2095 - 2102. [Abstract] [Full Text]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| INT J SYST EVOL MICROBIOL | MICROBIOLOGY | J GEN VIROL |
| J MED MICROBIOL | ALL SGM JOURNALS | |
