한국동물위생학회지 (Korean Journal of Veterinary Service)

  • 제43권4호
  • /
  • Pages.237-244
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  • 2020
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  • 3022-7372(pISSN)
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  • 2287-7630(eISSN)
한국동물위생학회 (The Korean Society of Veterinary Service)
권호 표지
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Development of sandwich enzyme-linked immunosorbent assay for a large-scale detection of porcine transmissible gastroenteritis virus in feces

  • Oh, Yeonsu (Department of Veterinary Pathology, Collage of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University) ;
  • Lee, Sang-Joon (Department of Veterinary Pathology, Collage of Veterinary Medicine and Institute of Veterinary Science, Kangwon National University) ;
  • Cho, Ho-Seong (College of Veterinary Medicine and Bio-Safety Research Institute, Jeonbuk National University) ;
  • Tark, Dongseob (Korea Zoonosis Research Institute, Jeonbuk National University)
  • 투고 : 2020.11.24
  • 심사 : 2020.12.17
  • 발행 : 2020.12.30
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초록

Porcine transmissible gastroenteritis (TGE) has been a significant cause of economic losses in pig farming industry since 1950s. Although transmissible gastroenteritis virus (TGEV) has declined in recent years, it should not be excluded because of its characteristics; the frequency of gene mutation, the mortality in piglets, and the possibility for sudden incidence. Therefore, the herd-level monitoring of the virus is important to prevent further circulation of TGE. The aim of this study is to develop a large-scale sandwich enzyme-linked immunosorbent assay (ELISA) with high specificity to rapidly detect TGEV in feces by using monoclonal antibodies (Mabs). The TGEV specific Mabs were produced in hybridoma cells. Among the Mabs belonged to the IgG class developed by this study, the final selected 8H6, 1B7, 4G3, and 1F8 were identified to have the neutralization ability against TGEV. The sandwich ELISA was established using 8H6 as a reporter antibody and 1B7 and the reported 5C8 as a capture antibody. The developed sandwich ELISA was able to distinguish TGEV from other pathogenic diarrheal agents (porcine rotavirus, porcine reovirus, porcine epidemic diarrhea virus (PEDV), E. coli, and C. perfringens) in tissue culture as well as fecal samples. And the detection rate of TGEV in feces was 80% compared with RT-PCR. The results suggested that the developed sandwich ELISA may be useful in the herd-level monitoring for effective preventive measures due to the early diagnosis of TGEV using a large amount of samples.

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참고문헌

  1. Asagi M, Ogawa T, Minetoma T, Sato K, Inaba Y. 1986. Detection of transmissible gastroenteritis virus in feces from pigs by reversed passive hemagglutination. Am J Vet Res 47(10): 2161-2164.
  2. Bernard S, Laude H, Lantier I. 1986. Detection du coronavirus de la GET et des anticorps specifiques par une technique ELISA mixte mettant en ouvre anticorps monoclonaux et policlonaux. J Rech Porcine Fr 18: 363-370.
  3. Black, JW. 1971. Diagnosis of TGE by FA: Evaluation of accuracy on field specimens. Proc US Anim Health Assoc 75: 492-498.
  4. Bohac J, Derbyshire JB, Thorsen J. 1975. The detection of transmissible gastrenteritis viral antigens by immunodiffusion. Can J Comp Med 39(1): 67-75.
  5. Chang YE, Cho SH, Kang SY. 1995. Monoclonal Antibody Production, Characterization and Utilization of porcine transmissible gastroenteritis virus. Korean Journal of Veterinary Medicine, 35 (4): 135.
  6. Chu RM, Li NJ, Glock RD, Ross RF. 1982. Applications of peroxidase-antiperoxidase staining technique for detection of transmissible gastroenteritis virus in pigs. Am J Vet Res 43(1): 77-81.
  7. Colye PV, Wyatt D, McCaughey C, O'Neil HJ. 1992. A simple standardised protocol for the production of monoclonal antibodies against viral and bacterial antigens. J Immunol Methods 153: 81-84. https://doi.org/10.1016/0022-1759(92)90308-G
  8. Dulac GC, Ruckerbauer GM, Boulanger P. 1977. Transmissible gastroenteritis: Demonstration of the virus from field specimens by means of cell culture and pig inoculation. Can J Comp Med 41: 357-363.
  9. Hsu TH, Liu HP, Chin CY, Wang C, Zhu WZ, Wu BL, Chang YC. 2018. Detection, sequence analysis, and antibody prevalence of porcine deltacoronavirus in Taiwan. Archives of virology 163(11): 3113-3117. https://doi.org/10.1007/s00705-018-3964-x
  10. Lanza I, Rubio P, Munoz M, Carmenes P. 1993. Comparison of a monoclonal antibody capture ELISA (MACELISA) to indirect ELISA and virus neutralization test for the serodiagnosis of transmissible gastroenteritis virus. J Vet Diagn Invest 5(1): 21-25. https://doi.org/10.1177/104063879300500106
  11. Maes RK, Haelterman EO. 1979. A seroepizootiological study of five viruses in a swine-evaluation station. Am J Vet Res 40: 1642-1645.
  12. Nakane PK, Kawaoi. 1974. Peroxidase-labeled antibody: Anew method of conjugation. J Histochem Cytochem 22: 1804-1091. https://doi.org/10.1177/22.12.1084
  13. Oh Y, Tark D. 2019. Development of monoclonal antibody capture ELISA for the detection of antibodies against transmissible gastroenteritis virus. Korean J Vet Serv 42(1): 9-15. https://doi.org/10.7853/kjvs.2019.42.1.9
  14. Pensaert MB, Haelterman EO, Burnstein T. 1970. Transmissible gastroenteritis of swine: Virus-intestinal cell interactions. I. Immunofluorescence, histopathology and virus production in the small intestine through the course of infection. Arch Gesamte Virusforsch 31: 321-334. https://doi.org/10.1007/BF01253767
  15. Saif LJ, Bohl EH, Kohler EM, Hughes JH. 1977. Immune electron microscopy of transmissible gastroenteritis virus and rotavirus (reovirus-like agent) of swine. Am J Vet Res 38(1): 13-20.
  16. Van Nieuwstadt AP, Cornelissen JB, Zetstra T. 1988. Comparison of two methods for detection of transmissible gastroenteritis virus in feces of pigs with experimentally induced infection. Am J Vet Res 49: 1836-1843.
  17. Yolken RH, Stopa PJ. 1979. Analysis of nonspecific reactions in enzyme-linked immunosorbent assay testing of human rotavirus. J Clin Microbiol 10: 703-707. https://doi.org/10.1128/jcm.10.5.703-707.1979
  18. Yolken RH. 1982. Enzyme immunoassays for the detection of infectious antigens in body fluids: current limitations and future prospects. Res Infect Dis 4: 35-68. https://doi.org/10.1093/clinids/4.1.35
  19. Zhang F, Luo S, Gu J, Li Z, Li K, Yuan W, Ye Y, Li H, Ding Z, Song D, Tang Y. 2019. Prevalence and phylogenetic analysis of porcine diarrhea associated viruses in southern China from 2012 to 2018. BMC veterinary research 15(1): 1-9. https://doi.org/10.1186/s12917-018-1758-8