Cite:  Polupan І.М., & Rudoi О.V. (2024). Vydilennia virusu skazu v kulturi klityn [Isolation of rabies virus in cell culture]. Veterynarna biotekhnolohiia – Veterinary biotechnology, 44, 103-108. https://doi.org/10.31073/vet_biotech44-09 [in Ukrainian].

POLUPAN І.М., email: This email address is being protected from spambots. You need JavaScript enabled to view it.

Institute of Veterinary Medicine of the NAAS

RUDOI О.V., email: This email address is being protected from spambots. You need JavaScript enabled to view it.

State Research Institute for Laboratory Diagnostics and Veterinary and Sanitary Expertise

ISOLATION OF RABIES VIRUS IN CELL CULTURE 

    Introduction. The main method of diagnosing rabies is the Direct Fluorescent Antibody test (DFA). In case of doubtfull results, the mouse innoculation test is performed. Despite the fact that the mouse innoculation test is a highly sensitive method of rabies diagnostic, it has significant limitations. An alternative to the mouse innoculation test for the isolation of the rabies virus from pathological material is the use of cell culture.

   The goal of the work. To evaluate the suitability of various cell cultures for isolation of the rabies virus.

   Materials and methods of research. To conduct comparative studies on the isolation of rabies virus from pathological material, three types of cell cultures BHK-21 C13 (АТСС ССL-10), Neuro-2a (АТСС CCL-131) and SK (saiga kidney) were used. 11 samples of the pathological material (brain) were used: 3 from cats, 4 from dogs, 2 from foxes and 2 from cattle. The infectivity of the pathological material was tested by performing the mouse innoculation test. The presence of the genetic material of the rabies virus in the pathological material was tested by RT-PCR.

   Results of research and discussion. In the cell culture BHK-21 C13, the rabies virus was isolated in eight of 11 pathological materials, which made 72.7%. Low sensitivity to street isolates of the rabies virus was determined in the cells SK, in which only two out of 11 (18.2%) street isolates of the rabies virus were isolated. Cell culture Neuro-2a showed an absolute sensitivity of 100% (11 out of 11 materials were isolated) compared to the results of the mouse innoculation test and RT-PCR.

   Conclusions and prospects for further research. According to the results of the research, it was determined that the optimal cell culture for isolating the rabies virus from pathological material is a cells of mouse neuroblastoma Neuro-2a (ATCC CCL-131).

Keywords: rabies, laboratory diagnosis, pathological material, cell culture, mouse innoculation test, virus isolation.

REFERENCES

1. Veterynarna medytsyna. Metody diahnostyky skazu [Veterinary medicine. Methods of diagnosis of rabies]: DSTU 7053:2009. (2009). Kyiv: Derzhspozhyvstandart Ukrainy [in Ukrainian].
2. Webster, W.A. (1987). A tissue culture infection test in routine rabies diagnosis. Canadian journal of veterinary research = Revue canadienne de recherche veterinaire, 51(3), 367-369.
3. Barrat, J., Barrat, M. J., Picard, M., & Aubert, M.F. (1988). Diagnostic de la rage sur culture cellulaire. Comparison des résultats de l'inoculation au neuroblastome murin et de l'inoculation a la souris [Diagnosis of rabies by cell culture]. Comparative immunology, microbiology and infectious diseases, 11(3-4), 207-214. https://doi.org/10.1016/0147-9571(88)90039-2.
4. Madhusudana, S.N., Sundaramoorthy, S., & Ullas, P.T. (2010). Utility of human embryonic kidney cell line HEK-293 for rapid isolation of fixed and street rabies viruses: comparison with Neuro-2a and BHK-21 cell lines. International journal of infectious diseases: IJID: official publication of the International Society for Infectious Diseases, 14(12), e1067-e1071. https://doi.org/10.1016/j.ijid.2010.07.004.
5. Zimmer, K., Wiegand, D., Manz, D., Frost, J.W., Reinacher, M., & Frese, K. (1990). Evaluation of five different methods for routine diagnosis of rabies. Zentralblatt fur Veterinarmedizin. Reihe B. Journal of veterinary medicine. Series B, 37(5), 392-400. https://doi.org/10.1111/j.1439-0450.1990.tb01074.x.
6. Ivanov, M.Yu. (2011). Epizootolohichna ta molekulyarno-biolohichna kharakterystyka vulychnykh izolyativ virusu skazu v Ukraini [Epizootological and molecular-biological characteristics of street isolates of the rabies virus in Ukraine]. Candidate’s thesis. Kyiv [in Ukrainian].
7. Corona, T.F., Böger, B., Rocha, T.C.D., Svoboda, W.K., & Gomes, E.C. (2018). Comparative analysis of Mouse Inoculation Test and Virus Isolation in Cell Culture for rabies diagnosis in animals of Parana, Brazil. Revista da Sociedade Brasileira de Medicina Tropical, 51(1), 39-43. https://doi.org/10.1590/0037-8682-0303-2017.
8. Dorward, W.J. (1974). Laboratory Techniques in Rabies – Third Edition. The Canadian Veterinary Journal, 15(9), xix.
9. Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. Chapter 3.1.18. Rabies (infection with rabies virus and other lyssaviruses). (2024). https://www.woah.org/fileadmin/Home/eng/Health_standards/tahm/3.01.18_RABIES.pdf.

 Download full text in PDF