Suzutani T, Saijo M, Nagamine M, Ogasawara M, Azuma M
Department of Microbiology, Asahikawa Medical College, Asahikawa, Japan.
J Clin Microbiol. 2000 May;38(5):1839-44. doi: 10.1128/JCM.38.5.1839-1844.2000.
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.
为监测对阿昔洛韦耐药的病毒，开发了一种针对单纯疱疹病毒（HSV）和水痘带状疱疹病毒（VZV）胸苷激酶（TK）基因的快速表型筛选方法。该方法可确定TK多肽的生化表型，其通过如下步骤由病毒DNA体外合成。每个样本病毒株的TK基因在T7启动子控制下通过PCR进行扩增和分离。PCR产物用T7 RNA聚合酶转录并在兔网织红细胞裂解物中翻译。使用该方法，无需病毒分离即可在不到2天的时间内确定编码HSV和VZV DNA的TK多肽的酶学特性和大小。该检测方法应是监测耐药病毒的有力工具，尤其是在病毒分离困难的情况下。
