Rapid phenotypic characterization method for herpes simplex virus and Varicella-Zoster virus thymidine kinases to screen for acyclovir-resistant viral infection.

Abstract:

: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.

journal_name

J Clin Microbiol

authors

Suzutani T,Saijo M,Nagamine M,Ogasawara M,Azuma M

doi

10.1128/JCM.38.5.1839-1844.2000

subject

Has Abstract

pub_date

2000-05-01 00:00:00

pages

1839-44

issue

5

eissn

0095-1137

issn

1098-660X

journal_volume

38

pub_type

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