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Journal of Clinical Microbiology, May 2000, p. 1839-1844, Vol. 38, No. 5
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Rapid Phenotypic Characterization Method for Herpes Simplex Virus and Varicella-Zoster Virus Thymidine Kinases To Screen for Acyclovir-Resistant Viral Infection

Tatsuo Suzutani,1,* Masayuki Saijo,2 Masayoshi Nagamine,1 Masahiro Ogasawara,1 and Masanobu Azuma1

Department of Microbiology, Asahikawa Medical College, Asahikawa,1 and Special Pathogens Laboratory, National Institute of Infectious Diseases, Tokyo,2 Japan

Received 1 October 1999/Returned for modification 5 January 2000/Accepted 21 February 2000

A rapid phenotypic screening method for herpes simplex virus (HSV) and varicella-zoster virus (VZV) thymidine kinase (TK) genes was developed for monitoring acyclovir-resistant viruses. This method determines the biochemical phenotype of the TK polypeptide, which is synthesized in vitro from viral DNA using a procedure as follows. The TK gene of each sample virus strain is amplified and isolated under the control of a T7 promoter by PCR. The PCR products are transcribed with T7 RNA polymerase and translated in a rabbit reticulocyte lysate. Using this method, enzymatic characteristics and the size of the TK polypeptides encoding HSV and VZV DNA were defined in less than 2 days without virus isolation. The assay should be a powerful tool in monitoring drug-resistant viruses, especially in cases in which virus isolation is difficult.

* Corresponding author. Mailing address: Department of Microbiology, Asahikawa Medical College, 2-1-1-1 Midorigaoka-Higashi, Asahikawa 078-8510, Japan. Phone: 81-166-68-2393. Fax: 81-166-68-2399. E-mail: suzutani{at}asahikawa-med.ac.jp.

Journal of Clinical Microbiology, May 2000, p. 1839-1844, Vol. 38, No. 5
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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