Korean Journal of Veterinary Research (대한수의학회지)

The Korean Society of Veterinary Science (대한수의학회)
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Optimalization of ELISA using Recombinant p27 Protein of SIV for Detection of Anti-SIV

SIV의 p27 재조합 단백질을 이용한 SIV 항체 검출을 위한 ELISA의 최적 조건

  • Kim, Eun-ok (College of Veterinary Medicine, Chungnam National University) ;
  • Kim, Eun (College of Veterinary Medicine, Chungnam National University) ;
  • Oh, Yoon-i (College of Veterinary Medicine, Chungnam National University) ;
  • Shin, Kwang-soon (College of Veterinary Medicine, Chungnam National University) ;
  • Kim, Hyun-soo (College of Veterinary Medicine, Chungnam National University) ;
  • Kim, Chul-joong (College of Veterinary Medicine, Chungnam National University)
  • 김은옥 (충남대학교 수의과대학) ;
  • 김은 (충남대학교 수의과대학) ;
  • 오윤이 (충남대학교 수의과대학) ;
  • 신광순 (충남대학교 수의과대학) ;
  • 김현수 (충남대학교 수의과대학) ;
  • 김철중 (충남대학교 수의과대학)
  • Accepted : 2002.01.27
  • Published : 2002.03.25
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Abstract

The p27 coding region of the SIVmac239 isolate was amplified by PCR and cloned into an expression vector, pMAL-cri, which expressed high levels of the p27 protein from Escherichia coli. The purified p27 protein was used for detection of anti-SIV antibodies with the sera from 11 macaques and 21 marmosets by immunoblot assay of which one macaque was suspicious for the SIV infection. The optimum conditions of ELISA was studied by the check board system with the recombinant purified p27 protein. For the plate coating, 200ng/well of the purified p27 was satisfactory. The conjugate was diluted 1:1000. The sera from the 32 monkeys were negative for the anti-SIV by ELISA.

SIVmac239 isolate의 p27유전자를 PCR로 증폭하여 pMAL-cri vector에 cloning하였다. 이를 Escherichia coli.에서 발현시켜 정제하여 국내에 수입된 macaque 11마리와 marmoset 21마리의 혈청에서 SIV 항체를 immunoblot으로 검사한 결과 macaque 1마리가 SIV 감염이 의심되었다. 또한 정제된 재조합 단백질을 이용하여 check board system으로 ELISA 방법의 최적 조건을 확립하였다. 정제 p27 항원 200ng/well을 plate에 coating하기에 적합하였고 conjugate는 1:1000으로 희석하는 것이 가장 좋았으며 위의 32 마리의 혈청을 ELISA로 검사한 결과 모두 anti-SIV 음성이었다.

Keywords

References

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