Optimized assay for thymidine kinase and its application to the detection of antibodies against herpes simplex virus type 1- and 2-induced thymidine kinase

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Abstract
An optimized assay for herpes simplex virus type 1- and 2-induced deoxythymidine kinase (dTk) is described which used [125I]iododeoxyuridine (IUdR) as a substrate. Values for Km and Vsat were determined for viral and cellular dTK, using either deoxythymidine or IUdR as a substrate. A comparison between the 2 substrates revealed that higher reaction velocities and lower Km values were obtained with IUdR. A standard assay was designed which uses 10-7 M IUdR as a substrate. This assay can detect herpes simplex-induced dTK from as few as 2 infected cells and is several orders of magnitude more sensitive than conventional dTK assays which use 10-5 M dT as a substrate. The assay was also used for detection of antibodies capable of blocking viral dTK. An easily detectable blocking activity, which was mainly confined to the immunoglobulin G antibody class, was found in most human sera which were positive for complement-fixing antibody against herpes simplex virus.

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