Thymidine plaque autoradiography of thymidine kinase-positive and thymidine kinase-negative herpes viruses

R. B. Tenser, J. C. Jones, S. J. Ressel, F. A. Fralish

Research output: Contribution to journalArticlepeer-review

37 Scopus citations

Abstract

Plaques formed by herpes simplex virus (HSV), pseudorabies virus, and varicella-zoster virus were studied by plaque autoradiography after [14C]thymidine labeling. Standard thymidine kinase-positive (TK+) viruses and TK- mutants of HSV types 1 and 2 and pseudorabies virus were studied, including cell-cultured viruses and viruses isolated from animals. Autoradiography was performed with X-ray film with an exposure time of 5 days. After development of films, TK+ plaques showed dark rims due to isotope incorporation, whereas TK- plaques were minimally labeled. Plaque autoradiography of stock TK- viruses showed reversion frequencies to the TK+ phenotype of <10-3. Autoradiography indicated that TK- virus retained the TK- phenotype after replication in vivo. In addition, it was shown that TK- HSV could be isolated from mouse trigeminal ganglion tissue after corneal inoculation of TK- HSV together with TK+ HSV. The plaque autoradiographic procedure was very useful to evaluate proportions of TK+ and TK- virus present in TK+-TK- virus mixtures.

Original languageEnglish (US)
Pages (from-to)122-127
Number of pages6
JournalJournal of clinical microbiology
Volume17
Issue number1
DOIs
StatePublished - 1983

All Science Journal Classification (ASJC) codes

  • Microbiology (medical)

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