한국어병학회지 (Journal of fish pathology)

  • 제22권2호
  • /
  • Pages.167-172
  • /
  • 2009
  • /
  • 1226-0819(pISSN)
  • /
  • 2233-5412(eISSN)
한국어병학회 (The Korean Society of Fish Pathology)
권호 표지

넙치 혈청의 보존 조건이 ELISA 결과에 미치는 영향

Effect of storage conditions of olive flounder Paralichthys olivaceus serum on enzyme-linked immunosorbent assay

  • 김위식 (전남대학교 식품.수산생명의학부) ;
  • 장민석 (전남대학교 식품.수산생명의학부) ;
  • 김종오 (전남대학교 식품.수산생명의학부) ;
  • 김두운 (전남대학교 식품.수산생명의학부) ;
  • 정성주 (전남대학교 식품.수산생명의학부) ;
  • 김석렬 (국립수산과학원 병리연구과) ;
  • 박명애 (국립수산과학원 병리연구과) ;
  • 오명주 (전남대학교 식품.수산생명의학부)
  • Kim, Wi-Sik (Division of Food Science and Aqualife Medicine, Chonnam National University) ;
  • Jang, Min-Seok (Division of Food Science and Aqualife Medicine, Chonnam National University) ;
  • Kim, Jong-Oh (Division of Food Science and Aqualife Medicine, Chonnam National University) ;
  • Kim, Du-Woon (Division of Food Science and Aqualife Medicine, Chonnam National University) ;
  • Jung, Sung-Ju (Division of Food Science and Aqualife Medicine, Chonnam National University) ;
  • Kim, Seok-Ryel (Pathology division, National Fisheries Research and Development Institute) ;
  • Park, Myoung-Ae (Pathology division, National Fisheries Research and Development Institute) ;
  • Oh, Myung-Joo (Pathology division, National Fisheries Research and Development Institute)
  • 발행 : 2009.08.01
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

본 연구에서는 넙치 혈청의 보존 조건이 ELISA 결과에 미치는 영향을 확인하기 위하여, 항 BSA 넙치 혈청을 이용해 보존 온도, 보존 기간 및 해동 횟수에 따른 ELISA값을 비교하였다. 넙치 혈청을 보존 온도 (-80, -20, 4 및 20${^{\circ}C}$) 및 기간 (1, 30, 69 및 124일)에 따른 ELISA OD 값을 비교한 결과, 모든 시료에서 뚜렷한 OD값의 차이는 확인되지 않았다. -80℃ 및 -20${^{\circ}C}$에 보존된 혈청을 사용하여 해동횟수 (1, 5, 10회)에 따른 ELISA OD값을 비교한 결과, 해동을 반복한 혈청 및 미동결 혈청의 OD값은 뚜렷한 차이를 보이지 않았다. 이상의 결과로 넙치 혈청은 -80, -20, 4 및 20${^{\circ}C}$에서 124일간 보존하거나 혈청을 10회 반복 해동하여도 ELISA 결과에 영향을 끼치지 않는 것이 확인되었다.

Antibody-detection enzyme-linked immunosorbent assay (ELISA) was performed to determine whether the absorbance value of ELISA is influenced by the different serum storage conditions of olive flounder Paralichthys olivaceus. Flounder antiserum to bovine serum albumin was stored at -80, -20, 4 and 20${^{\circ}C}$ for 1, 30, 69 and 124 days, respectively. In addition, the flounder antisera were frozen at -80 and -20${^{\circ}C}$, respectively and then repeated 1, 5 and 10 thaw-freeze cycles. No significant difference was shown in ELISA optical density (O.D.) values of sera, which were stored at the above mentioned storage conditions during 124 days. ELISA O.D. values of unfrozen serum samples, which were previously stored at 4${^{\circ}C}$, were almost similar to those of sera undergoing 1, 5, and 10 freeze-thaw cycles after stored at -80 or -20${^{\circ}C}$. In conclusion, the ELISA O.D. values of flounder sera were not affected by various storage conditions: different temperatures (-80, -20, 4 and 20${^{\circ}C}$), durations of storage (1, 30, 69 and 124 days), and repeated thaw-freeze cycles (1, 5, and 10 times).

키워드

참고문헌

  1. Corbel, M.J.: The immune response in fish: areview. J. Fish Biol., 7: 539-563,1975 https://doi.org/10.1111/j.1095-8649.1975.tb04630.x
  2. Crowther, J.R.: ELISA: Theory and practice, Humana Press, New Jersey, pp.1-223, 1995
  3. De Marco, M.A., Foni, E., Campitelli, L., Raffini, E., Delogu, M. and Donatelli, I.: Long-term monitoring for avian influenza viruses inwild bird species in Italy. Vet. Res. Commun.,27: 107-114, 2003 https://doi.org/10.1023/B:VERC.0000014126.72654.22
  4. Kim, W.S.: Establishment of enzyme-linked immunosorbent assay (ELISA) system to detect specific antibodies against infectious hematopoietic necrosis virus (IHNV). Ph. D thesis, Hokkaido University, Japan, 2008
  5. LaPatra, S.E.: The use of serological techniques for virus surveillance and certification of finfish. Annu. Rev. Fish Dis., 6: 15-28, 1996 https://doi.org/10.1016/S0959-8030(96)90003-4
  6. Nitzan, S., Shwartsburd, B. and Hellerb, E.D.: The effect of storing hybrid bass (Morone saxatilis x M. chrysops) sera at freezing temperatures on antibody titers. Fish Shellfish Immunol., 15: 341-347, 2003 https://doi.org/10.1016/S1050-4648(02)00179-1
  7. Renzullo, P.O., Sateren, W.B., Garner, R.P., Milazzo, M.J., Birx, D.L. and McNeil, J.G. : HIV- 1 seroconversion in united states army active duty personnel, 1985-1999. AIDS, 15: 1569-1574, 2001 https://doi.org/10.1097/00002030-200108170-00015
  8. Shi, Y., Yi, Y., Li, P., Kuang, T., Li, L., Dong, M., Ma, Q. and Cao, C.: Diagnosis of severe acute respiratory syndrome (SARS) coronavirus nucleocapsid antibodies in an antgencapturing enzyme-linked immunosorbent assay. J. Clin. Microbiol., 41: 5781-5782, 2003 https://doi.org/10.1128/JCM.41.12.5781-5782.2003
  9. Watanabe, K., Nishizawa, T. and Yoshimizu, M.:Selection of brood stock candidates of barfin flounder using an ELISA system with recombinant protein of barfin flounder nervous necrosis virus. Dis. Aquat. Org., 41: 219-223, 2000 https://doi.org/10.3354/dao041219
  10. Watanabe, K., Suzuki, S., Nishizawa, T., Suzuki, K., Yoshimizu, M. and Ezura, Y.: Control strategy for viral nervous necrosis of barfin flounder. Fish Pathol., 33: 445-446, 1998 https://doi.org/10.3147/jsfp.33.445
  11. Wilson, M.R. and Warr, G.W.: Fish immunoglobu-lins and the genes that encode them. Annu.Rev. Fish Dis., 201-221, 1992 https://doi.org/10.1016/0959-8030(92)90064-5
  12. 粉川愉記: コイヘルペスウイルス（KHV）に對す る抗體を檢出するためのELISA法に關す る硏究. 修士學位論文, 北海道大學, 日本, 2006
  13. 奧田律子: サクラマスのスモルト生産における細 菌性腎臟病原因菌の感染環の形成とその防除對策に關する硏究. 博士學位論文, 北海道大學, 日本, 2007