Influenza virus H5N2 cold-adapted vaccine strain reproduction in MDCK and Vero cell cultures using plant proteases and culture media based on hydrolyzates of soya and rice flour | Репродукция холодоадаптированного вакцинного штамма вируса гриппа H5N2 в клеточных культурах MDCK и Vero при использовании растительных протеаз и питательных сред на основе гидролизатов рисовой и соевой муки
2010
Mazurkova, N.A. | Ryabchikova, E.I. | Ternovoj, V.A. | Troshkova, G.P. | Shishkina, L.N., State Research Center of Virology and Biotechnology Vektor, Novosibirsk Region (Russian Federation) | Desheva, Yu.A. | Rudenko, L.G., Russian Academy of Medical Sciences, St. Petersburg (Russian Federation). Research and Development Inst. of Experimental Medicine
A possibility of animal-nature protease trypsin replacement by plant proteases bromelayn and papain in order to reproduce a cold-adapted reassortant vaccine strain of influenza virus (A/17/duck/Potsdam/86/92 (H5N2) in Vero and MDCK cell cultures has been examined. The cells were propagated using Axcevir and DMEM culture media or experimental media based on enzymatic soy and rice flour hydrolysates. The yield of A/17/duck/Potsdam/86/92(H5N2) virus propagated in MDCK and Vero cells at the addition of papain reached 10E8.5-8.6 and 10E7.1-8.1 TCD50/ml; respectively; the value obtained using the experimental bromelayn medium was 10E9.3-9.4 and 10E7.9-8.5 TCD50/ml, respectively. The cold-adapted reassortant strain propagated in the experimental culture media in the presence of plant proteases retained phenotype and genetic features identical to those of the original vaccine strain. Ultrastructural parameters of the virus reproduction were also unaltered under different experimental conditions. Thus, using plant enzymes and media based on enzyme plant hydrolysates for cultivation of the vaccine strain on MDCK and Vero cell cultures allows ecologically safe culture vaccines against influenza virus
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