EXPERIENCE IN DECONTAMINATION OF VIRAL STRAINS FROM MYCOPLASMAS

Mycoplasma contamination is a serious problem for the production of biologics in the veterinary and biopharmacological industries, as well as for research laboratories. A number of dificulties are associated with the detection and decontamination of mycoplasma due to its high resistance to antibiotics and the latent nature of the course of mycoplasma infection. The development of effective methods for the timely detection of mycoplasmas and the decontamination of viral strains passaged in cell cultures is an urgent task. The aim of this work was to assess the possibility of decontamination of viruses from mycoplasma contamination using antibiotics. We used a collection of strains of orthohantaviruses (FSCIRIP named after M. P. Chumakov RAS) Puumala virus, Hantaan virus, Sochi virus - a genovariant of the Dobrava/Belgrade virus. A cytochemical method using Hoechst 33258 dye was used to indicate Mycoplasma. Virus reproduction was controlled by indirect method of fluorescent antibodies, the virus titer in the cell was determined by the method of focus-forming units. This paper presents data on Mycoplasma decontamination of orthohantavirus strains using the antibiotics ciprofloxacin and BMCyclin (Roche). It has been shown that ciprofloxacin has a clear cytotoxic effect, and the use of BMCyclin can lead to suppression of virus replication in the cell. A method of using BMCyclin for Hantaan orthohantavirus is proposed, which provides good survival of Vero E6 cell culture and a high degree of virus reproduction. The high eficiency of the antibiotic BMCyclin for decontamination of Mycoplasma has been shown. Its use is simple, effective and low-cost and can be recommended in order to preserve valuable viral strains.

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