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

Infection and immunity ·Vol. 29 ·No. 2 ·1980-08-00 ·Pages 425-34

Gronowitz JS, Källander CF

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 both viral and cellular dTK, using either deoxythymidine or IUdR as a substrate. A comparison between the two 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 two infected cells and is several orders of magnitude more sensitive than conventional dTK assays which use 10(-5) M dT as a substrate. An easily detectable blocking activity, which was shown to be 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.

MeSH Terms

Antibodies, Viral/analysis Electrophoresis, Polyacrylamide Gel/methods Humans Idoxuridine/metabolism Immunoglobulin G Simplexvirus/enzymology,immunology Thymidine Kinase/analysis

Chemicals

Antibodies, Viral Immunoglobulin G Thymidine Kinase Idoxuridine

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Gronowitz J S

Källander C F

References (23)

23 references, click to expand

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Article Info

Journal

Infection and immunity

Abbr.

Infect Immun

ISSN

0019-9567

DOI

10.1128/iai.29.2.425-434.1980

Published

1980-08-00

Pages

425-34

Language

English

Region

United States

NLM ID

0246127

PMCID

PMC551136

Subset

External Links

PubMed source DOI: 10.1128/iai.29.2.425-434.1980 PMC Full Text View journal details

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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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