1. Porcine retrovirus: optimal conditions for its biochemical detection.
- Author
-
Phan-Thanh L, Kaeffer B, and Bottreau E
- Subjects
- Animals, Cell Line, Detergents pharmacology, Hydrogen-Ion Concentration, Magnesium pharmacology, Male, Manganese pharmacology, Microscopy, Electron, Potassium pharmacology, RNA, Messenger metabolism, Retroviridae enzymology, Retroviridae ultrastructure, Retroviridae Infections microbiology, Retroviridae Infections transmission, Sodium pharmacology, Swine, Templates, Genetic, Transcription, Genetic, RNA-Directed DNA Polymerase metabolism, Retroviridae isolation & purification
- Abstract
The assay of reverse transcriptase (RT) activity was used to detect the presence of retrovirus in porcine cells. A set of optimal assay conditions was determined to design a sensitive, quantitative and reproducible RT assay for porcine systems. The template-primer poly(rA).oligo(dT) was an absolute requirement. The presence of Mn++ was indispensable, with an optimal concentration of 0.25 mM. Monocations (K+, Na+) at 50 mM greatly enhanced, but their high doses inhibited the reaction. The pH of the medium influenced very much the reaction, especially with non-purified virus samples, with which the RT activity was inhibited at pHs above 8.2. Non-ionic detergents at 1% enhanced several-fold the RT activity. It was also shown that porcine retrovirus could be spontaneously reactivated in porcine cell lines by in vitro long-term propagation and transmitted to pigs by inoculation with virus-producing cells.
- Published
- 1992
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