Development of ELISA for Brucella abortus RB51 II. Purification of 8kDa antigen and development of ELISA using its antigen of Brucella abortus RB51

부루세라 RB51의 ELISA 진단법 개발 II. Brucella abortus RB51균의 8kDa 항원 정제 및 ELISA 진단법 개발

  • Her, Moon (Bacteriology and Parasitology Division, National Veterinary Research and Quarantine Services) ;
  • Cho, Dong-hee (Bacteriology and Parasitology Division, National Veterinary Research and Quarantine Services) ;
  • Jung, Byeong-yeal (Bacteriology and Parasitology Division, National Veterinary Research and Quarantine Services) ;
  • Cho, Seong-kun (Bacteriology and Parasitology Division, National Veterinary Research and Quarantine Services) ;
  • Jung, Suk-chan (Bacteriology and Parasitology Division, National Veterinary Research and Quarantine Services) ;
  • Kim, Ok-kyung (Bacteriology and Parasitology Division, National Veterinary Research and Quarantine Services)
  • 허문 (국립수의과학검역원 세균과) ;
  • 조동희 (국립수의과학검역원 세균과) ;
  • 정병열 (국립수의과학검역원 세균과) ;
  • 조성근 (국립수의과학검역원 세균과) ;
  • 정석찬 (국립수의과학검역원 세균과) ;
  • 김옥경 (국립수의과학검역원 세균과)
  • Accepted : 2000.12.04
  • Published : 2001.03.20

Abstract

A procedure for extraction and purification of 8 kDa antigen of Brucella abortus RB51 was developed. Bacteria heat inactivated at $60^{\circ}C$, 30 min was extracted by 1% sarcosine and followed by fluid pressure liquid gel filtration chromatography of 2 series, Superose 12 HR 10/30 and Sephacryl S-100. There was produced $71.46{\mu}g/g$(wet) of 8 kDa antigen, and it resisted 1% trypsin, solved 1% triton X-100 higher than distilled water and inactivated 0.1% proteinase K. These results show that 8 kDa antigen may be a lipoprotein existed cell surface of B. abortus RB51. Also, we developed ELISA using purified 8 kDa surface antigen of Brucella abortus RB51 strain, its specificity and sensitivity was 95.0%, 98.6%, respectively. As compared with dot-blot assay using whole cell and ELISA using 8 kDa antigen, its correlation was 93.5%.