Storage Methods of Complex RNA Viruses

Viktoriya V. Varianytsia, Igor P. Vysekantsev


The review describes the development of methods for the long-term storage of complex RNA viruses. The need of such an activity is stipulated by a wide spreading of infectious diseases caused by these viruses and rising number of the studies dealing with developing the drugs for immune prophylaxis and therapy. Among other things the technology for these drugs production involves the long-term storage of viruses. This includes freeze-drying and storage at temperatures below0°C. However, to date no generally accepted approaches exist in terms of composition of protective media for either freeze-drying or low-temperature storage of viruses. For this purpose, one uses the numerous additives and combinations of various sugars, polyhydric alcohols, animal proteins, chemical compounds. Storage of frozen-dried viruses is performed at either 4 or –20°C, and in case of a frozen state it ranges from –60 to –85°C, that is stipulated by the availability of freezing chambers providing these temperatures, and high survival of viruses from different families.

Probl Cryobiol Cryomed 2017; 27(4): 287–295


complexly constructed RNA-containing viruses; long-term storage; freeze-drying; stabilizing substances; protective media; virus preservation rate; vaccines

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ATCC Virology guide. Tips and techniques for propagating virus in tissue culture and embryonated chicken eggs. Manassas: ATCC; 2016.

Bean B., Moore B.M., Sterner B. et al. Survival of influenza viruses on environmental surfaces. Journal of Infectious Diseases 1982; 146(1): 47–51. CrossRef PubMed

Brovarnik V.V., Golovach T.M. Information support of the microorganisms cultures depository. Control Systems and Machines 2015; (4): 58-66.

Burkova V.V., Visekantsev I.P., Lavrik A.A. Preservation of infectious activity of rabies virus industrial strains stored at various temperatures // Electronic periodical publication of SFU 'Live and bioconcent systems'. – 2014; (9). – Available from: -9/article – 2.

Carpenter J.F., Izutsu K., Randolph T.W. Freezing- and dryinginduced perturbations of protein structure and mechanisms of protein protection by stabilizing additives. In: L. Rey, J.C. May, editors. Freeze Drying/Lyophilisation of Pharmaceutical and Biological Products. 3d ed. London: Informa Healthcare; 2010: 167–197. PubMed

Chaniot S.C.M., Holmes M.J., Stott E.J., Tyrrell D.A.J. An investigation of media for the long term storage of three respiratory viruses. Arch Virol 1974; 44(4): 396–400. CrossRef

Fernie B.F., Gerin J.L. The stabilization and purification of respiratory syncytial virus using MgSO4. Virology 1980; 106(1): 141–144. CrossRef

Gould E.A. Methods for long-term virus preservation. Mol Biotechnol 1999; 13(1): 57–66. CrossRef

Greiff D. Stabilities of suspensions of influenza virus dried by sublimation of ice in vacuo to different contents of residual moisture and sealed under different gases. Appl Microbiol 1970; 20(6): 935–938. PubMed

Greiff D., Rightsel W. A. Stabilities of suspensions of viruses after freezing or drying by vacuum sublimation and storage. Cryobiology 1967; 3(6): 432–444. CrossRef

Gupta C.K., Leszczynski J., Gupta R.K., Siber G.R. Stabilization of respiratory syncytial virus (RSV) against thermal inactivation and freeze-thaw cycles for development and control of RSV vaccines and immune globulin. Vaccine 1996; 18(15): 1417–1420. CrossRef

Howell C.L., Miller M.J. Effect of sucrose phosphate and sorbitol on infectivity of enveloped viruses during storage. J Clin Microbiol 1983; 18(3): 658–662.

Hubalek Z. Protectants used in the cryopreservation of microorganisms. Cryobiology 2003; 46(3): 205–229. CrossRef

Johnson F.B. Transport of viral specimens. Clinical Microbiology Reviews 1990; 3(2): 120–131. CrossRef

Lvov D.K., Alimbarova L.M., Alkhovsky S.V. et al. Medical virology. Moscow: MIA; 2008.

Malenovska H. The influence of stabilizers and rates of freezing on preserving of structurally different animal viruses during lyophilization and subsequent storage. J Appl Microbiol 2014; 117(6): 1810–1819. CrossRef PubMed

McAleer W.J., Markus H.Z., McLean A. A. et al. Stability on storage at various temperatures of live measles, mumps and rubella virus vaccines in new stabilizer. Journal of Biological Standardization 1980: 8(4): 281–287. CrossRef

Meslin F.X., Kaplan M.M., Koprowski H. Laboratory techniques in rabies. Geneva: World Health Organization; 1996.

Pisal S., Wawde G., Salvankar S. et al. Vacuum foam drying for preservation of LaSota virus: effect of additives. AAPS PharmSciTech 2006; 7(3): E30–E37. CrossRef PubMed

Polupan I.M. Estimation of different storage methods of rabies virus vaccine strains. Veterinary Biotechnology 2011; (20): 127–133.

Rabies. In: OIE terrestrial manual. Paris: World Organisation for Animal Health; 2013. p. 1–28.

Sarkar J., Sreenivasa B.P., Singh R.P. et al. Comparative efficacy of various chemical stabilizers on the thermostability of a liveattenuated peste des petits ruminants (PPR) vaccine. Vaccine 2003; 21(32): 4728–4735. CrossRef

Stegniy M.Yu. Properties of cattle viruses, preserved at moderately low temperatures. Probl Cryobiol 2003; (3): 88–91.

Stroganova I.Ya. Infectious activity of cattle respiratory syncytial virus during storage. Bulletin of Krasnoyarsk State Agrarian University 2009; (12): 155–157.

Tanaka T., Takeda T., Miyajama R. Cryoprotective effect of saccharides on denaturation of catalase during freeze-drying. Chem Pharm Bull 1991; 675(5): 1091–1094. CrossRef

Tannock G.A., Hierholzer J.C., Bryce D.A. et al. Freeze-drying of respiratory syncytial viruses for transportation and storage. J Clin Microbiol 1987; 25(9): 1769–1771. PubMed

Tedeschi R., De Paoli P. Collection and preservation of frozen microorganisms. Methods Mol Biol 2011; 675: 313–326. CrossRef


Wallis C., Melnick J.L. Stabilization of enveloped viruses by dimethyl sulfoxide. J Virol 1968; 2(9): 953–954. PubMed

World Health Organization. Recommendations for inactivated rabies vaccine for human use produced in cell substrates and embryonated eggs. In: WHO Expert Committee on Biological Standardization. Fifty-sixth report. Geneva: World Health Organization; 2007. p. 83–132.



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