Journal of Veterinary Science

  • 제22권4호
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  • Pages.56.1-56.10
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  • 2021
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  • 1229-845X(pISSN)
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  • 1976-555X(eISSN)
대한수의학회 (The Korean Society of Veterinary Science)
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Generation of a recombinant rabies virus expressing green fluorescent protein for a virus neutralization antibody assay

  • Yang, Dong-Kun (Viral Disease Division, Animal and Plant Quarantine Agency, Ministry of Agriculture, Food and Rural Affairs) ;
  • Kim, Ha-Hyun (Viral Disease Division, Animal and Plant Quarantine Agency, Ministry of Agriculture, Food and Rural Affairs) ;
  • Park, Yu-Ri (Viral Disease Division, Animal and Plant Quarantine Agency, Ministry of Agriculture, Food and Rural Affairs) ;
  • Yoo, Jae Young (Viral Disease Division, Animal and Plant Quarantine Agency, Ministry of Agriculture, Food and Rural Affairs) ;
  • Park, Yeseul (Viral Disease Division, Animal and Plant Quarantine Agency, Ministry of Agriculture, Food and Rural Affairs) ;
  • Park, Jungwon (Viral Disease Division, Animal and Plant Quarantine Agency, Ministry of Agriculture, Food and Rural Affairs) ;
  • Hyun, Bang-Hun (Viral Disease Division, Animal and Plant Quarantine Agency, Ministry of Agriculture, Food and Rural Affairs)
  • 투고 : 2021.02.16
  • 심사 : 2021.06.29
  • 발행 : 2021.07.31
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초록

Background: Fluorescent antibody virus neutralization (FAVN) test is a standard assay for quantifying rabies virus-neutralizing antibody (VNA) in serum. However, a safer rabies virus (RABV) should be used in the FAVN assay. There is a need for a new method that is economical and time-saving by eliminating the immunostaining step. Objectives: We aimed to improve the traditional FAVN method by rescuing and characterizing a new recombinant RABV expressing green fluorescent protein (GFP). Methods: A new recombinant RABV expressing GFP designated as ERAGS-GFP was rescued using a reverse genetic system. Immuno-fluorescence assay, peroxidase-linked assay, electron microscopy and reverse transcription polymerase chain reaction were performed to confirm the recombinant ERAGS-GFP virus as a RABV expressing the GFP gene. The safety of ERAGS-GFP was evaluated in 4-week-old mice. The rabies VNA titers were measured and compared with conventional FAVN and FAVN-GFP tests using VERO cells. Results: The virus propagated in VERO cells was confirmed as RABV expressing GFP. The ERAGS-GFP showed the highest titer (108.0 TCID50/mL) in VERO cells at 5 days post-inoculation, and GFP expression persisted until passage 30. The body weight of 4-week-old mice inoculated intracranially with ERAGS-GFP continued to increase and the survival rate was 100%. In 62 dog sera, the FAVN-GFP result was significantly correlated with that of conventional FAVN (r = 0.95). Conclusions: We constructed ERAGS-GFP, which could replace the challenge virus standard-11 strain used in FAVN test.

키워드

과제정보

This work was supported financially by a grant (B1543083-2018-20-02) from Animal, and Plant Quarantine Agency, Ministry of Agriculture, Food and Rural Affairs (MAFRA), Republic of Korea.

참고문헌

  1. Dietzschold B, Schnell M, Koprowski H. Pathogenesis of rabies. Curr Top Microbiol Immunol. 2005;292:45-56.
  2. Li J, Ertel A, Portocarrero C, Barkhouse DA, Dietzschold B, Hooper DC, et al. Postexposure treatment with the live-attenuated rabies virus (RV) vaccine TriGAS triggers the clearance of wild-type RV from the Central Nervous System (CNS) through the rapid induction of genes relevant to adaptive immunity in CNS tissues. J Virol. 2012;86(6):3200-3210. https://doi.org/10.1128/JVI.06699-11
  3. Ondrejkova A, Suli J, Ondrejka R, Benisek Z, Franka R, Svrcek S, et al. Comparison of the detection and quantification of rabies antibodies in canine sera. Vet Med (Praha). 2012;47(8):218-221. https://doi.org/10.17221/5827-vetmed
  4. Servat A, Wasniewski M, Cliquet F. Tools for rabies serology to monitor the effectiveness of rabies vaccination in domestic and wild carnivores. Dev Biol (Basel). 2006;125:91-94.
  5. Yang DK, Kim HH, Cho IS. Strategies to maintain Korea's animal rabies non-occurrence status. Clin Exp Vaccine Res. 2018;7(2):87-92. https://doi.org/10.7774/cevr.2018.7.2.87
  6. World Organization of Animal Health (OIE). Terrestrial animal health code. 29th ed. Paris (France): OIE; 2019, Chapter 8.14: Infection with rabies; 570-575.
  7. Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. Rabies (Infection with Rabies Virus and Other Lyssaviruses) [Internet]. Paris: OIE; https://www.oie.int/fileadmin/Home/eng/Health_standards/tahm/3.01.17_RABIES.pdf. Updated 2019. Accessed 2020 Jan 2.
  8. Hu Q, Chen W, Huang K, Baron MD, Bu Z. Rescue of recombinant peste des petits ruminants virus: creation of a GFP-expressing virus and application in rapid virus neutralization test. Vet Res (Faisalabad). 2012;43(1):48. https://doi.org/10.1186/1297-9716-43-48
  9. Luo J, Zhao J, Tian Q, Mo W, Wang Y, Chen H, et al. A recombinant rabies virus carrying GFP between N and P affects viral transcription in vitro. Virus Genes. 2016;52(3):379-387. https://doi.org/10.1007/s11262-016-1313-2
  10. Burgado J, Greenberg L, Niezgoda M, Kumar A, Olson V, Wu X, et al. A high throughput neutralization test based on GFP expression by recombinant rabies virus. PLoS Negl Trop Dis. 2018;12(12):e0007011. https://doi.org/10.1371/journal.pntd.0007011
  11. Khawplod P, Inoue K, Shoji Y, Wilde H, Ubol S, Nishizono A, et al. A novel rapid fluorescent focus inhibition test for rabies virus using a recombinant rabies virus visualizing a green fluorescent protein. J Virol Methods. 2005;125(1):35-40. https://doi.org/10.1016/j.jviromet.2004.12.003
  12. Qin S, Volokhov D, Rodionova E, Wirblich C, Schnell MJ, Chizhikov V, et al. A new recombinant rabies virus expressing a green fluorescent protein: a novel and fast approach to quantify virus neutralizing antibodies. Biologicals. 2019;59:56-61. https://doi.org/10.1016/j.biologicals.2019.03.002
  13. Tang HB, Lu ZL, Wei XK, Zhong YZ, Zhong TZ, Pan Y, et al. A recombinant rabies virus expressing a phosphoprotein-eGFP fusion is rescued and applied to the rapid virus neutralization antibody assay. J Virol Methods. 2015;219:75-83. https://doi.org/10.1016/j.jviromet.2015.03.022
  14. Xue X, Zheng X, Liang H, Feng N, Zhao Y, Gao Y, et al. Generation of recombinant rabies virus CVS-11 expressing eGFP applied to the rapid virus neutralization test. Viruses. 2014;6(4):1578-1589. https://doi.org/10.3390/v6041578
  15. Yang DK, Kim HH, Lee SH, Jeong WH, Tark D, Cho IS. A genetically modified rabies vaccine (ERAGS) induces protective immunity in dogs and cattle. Clin Exp Vaccine Res. 2017;6(2):128-134. https://doi.org/10.7774/cevr.2017.6.2.128
  16. Wera E, Mourits MC, Siko MM, Hogeveen H. Cost-effectiveness of mass dog vaccination campaigns against rabies in Flores Island, Indonesia. Transbound Emerg Dis. 2017;64(6):1918-1928. https://doi.org/10.1111/tbed.12590
  17. Kim CH, Lee CG, Yoon HC, Nam HM, Park CK, Lee JC, et al. Rabies, an emerging disease in Korea. J Vet Med B Infect Dis Vet Public Health. 2006;53(3):111-115. https://doi.org/10.1111/j.1439-0450.2006.00928.x
  18. Kim JH, Hwang EK, Sohn HJ, Kim DY, So BJ, Jean YH. Epidemiological characteristics of rabies in South Korea from 1993 to 2001. Vet Rec. 2005;157(2):53-56. https://doi.org/10.1136/vr.157.2.53
  19. Kasempimolporn S, Sichanasai B, Saengseesom W, Puempumpanich S, Sitprija V. Stray dogs in Bangkok, Thailand: rabies virus infection and rabies antibody prevalence. Dev Biol (Basel). 2008;131:137-143.
  20. Have P, Alban L, Berndtsson LT, Cliquet F, Hostnik P, Rodeia SC, et al. Risk of rabies introduction by non-commercial movement of pets. Dev Biol (Basel). 2008;131:177-185.
  21. Patel AC, Upmanyu V, Ramasamy S, Gupta PK, Singh R, Singh RP. Molecular and immunogenic characterization of BHK-21 cell line adapted CVS-11 strain of rabies virus and future prospect in vaccination strategy. Virusdisease. 2015;26(4):288-296. https://doi.org/10.1007/s13337-015-0285-5