Källander C F, Gronowitz J S, Olding-Stenkvist E
J Clin Microbiol. 1983 Feb;17(2):280-7. doi: 10.1128/jcm.17.2.280-287.1983.
A sensitive enzyme assay utilizing [125I]iododeoxyuridine as the substrate and CTP as the phosphate donor in combination with isozyme-specific antisera was used for direct detection and typing of herpesvirus deoxythymidine kinase (dTk) in clinical specimens. An investigation of 16 coded vesicle fluid specimens, taken in connection with varicella-zoster virus (VZV) and herpes simplex virus infections, revealed viral dTk activity in 14 samples. All positive samples except one were taken within 5 days after the onset of illness. Serological typing of the dTk activities easily established whether the vesicles were caused by VZV, herpes simplex virus type 1, or herpes simplex virus type 2. The results were obtainable within 5 h and were in agreement with the results achieved by immunofluorescence tests or by virus isolation when positive. Acute- and convalescent-phase sera from patients with VZV infections were analyzed with regard to dTk isozyme composition. All sera collected within 5 days after the onset of varicella were found to contain elevated levels of dTk activity. By the use of isozyme-specific antisera and gel electrophoresis, it was possible to show the presence of both cellular and VZV dTk's. Among the 13 acute-phase sera from zoster patients, only 2 were found to be VZV dTk positive. Convalescent sera, in most cases collected 15 days or more after the onset of illness, were also found to be devoid of VZV dTk. The relevance of the results and the possible use of these methods for viral diagnostics are discussed.
一种灵敏的酶分析方法,利用[125I]碘脱氧尿苷作为底物,CTP作为磷酸供体,并结合同工酶特异性抗血清,用于直接检测临床标本中的疱疹病毒脱氧胸苷激酶(dTk)并进行分型。对16份与水痘-带状疱疹病毒(VZV)和单纯疱疹病毒感染相关的编码水疱液标本进行的调查显示,14份样本中存在病毒dTk活性。除一份样本外,所有阳性样本均在发病后5天内采集。dTk活性的血清学分型可轻松确定水疱是由VZV、单纯疱疹病毒1型还是单纯疱疹病毒2型引起的。结果可在5小时内获得,且与免疫荧光试验或病毒分离阳性时的结果一致。对VZV感染患者的急性期和恢复期血清进行了dTk同工酶组成分析。发现水痘发病后5天内采集的所有血清中dTk活性水平均升高。通过使用同工酶特异性抗血清和凝胶电泳,有可能显示细胞dTk和VZV dTk的存在。在13份带状疱疹患者的急性期血清中,仅发现2份为VZV dTk阳性。恢复期血清,大多在发病后15天或更长时间采集,也被发现不含VZV dTk。讨论了这些结果的相关性以及这些方法在病毒诊断中的可能用途。