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Porcine retrovirus: optimal conditions for its biochemical detection

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.
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Submitted on : Monday, June 1, 2020 - 6:43:56 PM
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Luu Phan Thanh, Bertrand Kaeffer, Elisabeth Bottreau. Porcine retrovirus: optimal conditions for its biochemical detection. Archives of Virology, Springer Verlag, 1992, 123 (3-4), pp.255-265. ⟨10.1007/BF01317262⟩. ⟨hal-02713180⟩



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