Korean Journal of Fisheries and Aquatic Sciences (한국수산과학회지)

The Korean Society of Fisheries and Aquatic Science (한국수산과학회)
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Detection of Specific Antibodies Against Viral Hemorrhagic Septicemia Virus in Infected Olive Flounder Paralichthys olivaceus Using Enzyme-Linked Immunosorbent Assay

Enzyme-linked immunosorbent assay를 이용한 바이러스성 출혈성 패혈증 바이러스 감염 넙치(Paralichthys olivaceus)의 특이 항체반응 검사

  • Hwang, Jee Youn (Aquatic Disease Control Division, National Institute of Fisheries Science) ;
  • Jang, Jin Hyeon (Aquatic Disease Control Division, National Institute of Fisheries Science) ;
  • Kim, Dong Jun (Aquatic Disease Control Division, National Institute of Fisheries Science) ;
  • Kwon, Mun Gyeong (Aquatic Disease Control Division, National Institute of Fisheries Science) ;
  • Seo, Jung Soo (Aquatic Disease Control Division, National Institute of Fisheries Science) ;
  • Hwang, Seong Don (Aquatic Disease Control Division, National Institute of Fisheries Science) ;
  • Son, Maeng-Hyun (Aquatic Disease Control Division, National Institute of Fisheries Science)
  • 황지연 (국립수산과학원 수산방역과) ;
  • 장진현 (국립수산과학원 수산방역과) ;
  • 김동준 (국립수산과학원 수산방역과) ;
  • 권문경 (국립수산과학원 수산방역과) ;
  • 서정수 (국립수산과학원 수산방역과) ;
  • 황성돈 (국립수산과학원 수산방역과) ;
  • 손맹현 (국립수산과학원 수산방역과)
  • Received : 2017.09.21
  • Accepted : 2017.10.24
  • Published : 2017.10.31
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Abstract

The viral hemorrhagic septicemia virus (VHSV) has an extensive host range, and infects farmed and wild fish inhabiting both freshwater and marine ecosystems. Enzyme-linked immunosorbent assay (ELISA) is highly useful in diagnosing viral hemorrhagic septicemia. However, ELISA shows high, non-specific background reaction with fish antibodies. In this study, we optimized the antigen and antibody concentrations used for detecting specific antibodies in VHSV-infected olive flounder to reduce non-specific binding, and improve the sensitivity of ELISA. The results suggested that OD (optical Density) values were valid when ELISA was performed with $0.1{\mu}g/well$ of virus, involving blocking with blocking buffer (Roth, Roti-Block), 1:300-1:600 dilution with flounder antisera, and 1:1000 dilution with anti-flounder IgM and HRP-conjugated goat anti-mouse IgG for detecting the VHSV antibody in flounder sera. Furthermore, 11 different VHSV strains isolated in Korea from 2012 to 2016 were used to infect the fish. The results showed no correlation between viral pathogenicity and antibody production. This research is a basic study on the application of antibody detection in the diagnosis of viral hemorrhagic septicemia in the olive flounder.

Keywords

References

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