Radiometric Methods for Rapid Diagnosis of Viral Infection.
Abstract
Two radiometric techniques were investigated for monitoring the effect of herpes simplex virus on the Deoxyribonucleic acid synthesis of monolayers of human embryonic lung fibroblasts. DNA synthesis of infected and uninfested cells was monitored by 3H-thymidine incorporation measured by liquid scintillation counting or by 125I-iododeoxyuridine (125I-IdU) incorporation measured non-destructively by gamma scintillation counting. Radiometric results were compared to those obtained from visual examination for cytopathic effects in the same cell line. Herpes simplex virus type-2 (HSV-2, 10 to the 6.8th power TCID50) infected cells showed a marked increase in 3H-thymidine incorporation 2-6 hr after infection. HSV-2 and herpes simplex type 1 (HSV-1) exhibited similar levels of sensitivity with increased incorporation being observed 72 hr after infection with 10 virions. The 3H-thymidine technique was utilized to assay HSV-1 infected mouse brains. Increased 125I-IdU incorporation was observed 6 hr after infection with 10 to the 5th power to 10 to 6.8th power HSV-1 virions, 24 hr with 10,000 virions, 48 hr with 1000 virions, and 72 hr with 10-100 virions. The increased 125I-IdU incorporation was completely inhibited by preneutralization with immune serum. These radiometric techniques for detection of viral effect on cellular metabolism are simple, objective, and quantitive.
Document Details
- Document Type
- Technical Report
- Publication Date
- Nov 22, 1976
- Accession Number
- ADA158046
Entities
People
- H. N. Wagner Jr.
- M. F. Tsan
Organizations
- Johns Hopkins University