Development of ELISA for Brucella abortus RB51 II. Purification of 8kDa antigen and development of ELISA using its antigen of Brucella abortus RB51 |
Moon Her, Dong-hee Cho, Byeong-yeal Jung, Seong-kun Cho, Suk-chan Jung, Ok-kyung Kim |
Bacteriology and Parasitology Division, National Veterinary Research and Quarantine Services |
부루세라 RB51의 ELISA 진단법 개발 II. Brucella abortus RB51균의 8kDa 항원 정제 및 ELISA 진단법 개발 |
허문, 조동희, 정병열, 조성근, 정석찬, 김옥경 |
국립수의과학검역원 세균과 |
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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%. |
Key Words:
Brucella abortus, RB51, 8kDa antigen, lipoprotein, ELISA |
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