Method for identifying i(mycobacterium) species and diagnosing reagent kit therefor
A technology for mycobacteria and mycobacterial antigens, which can be used in biochemical equipment and methods, analytical materials, and microbial determination/inspection, etc., and can solve problems such as the specificity of protein antigens.
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[0034] Preparation of KP90 / KS90
[0035] KP90 is prepared with starting materials derived from crude mycobacterial substance, as described in WO-A-94 / 14069 (the content of this invention is incorporated herein by reference). The starting material prepared with 1 gram of freeze-dried bacteria was centrifuged at 90,000×g at 4°C for 2 hours. The pellet was washed twice with PBS. Between washing steps, the samples were sonicated on ice for 6×30 seconds with 10 seconds interruption each time, and then centrifuged at 90,000×g, 4°C for 2 hours. Collect the pellet and resuspend it in 10 ml of 0.05M Tris buffer (TBS) pH 7.4. The supernatant designated as KS90 can be used as an antigen. The precipitate was sonicated on ice for 6×15 seconds, with 10 seconds interruption each time. This formulation is designated the term KP90. After determining the protein concentration, 1.7 ml was lyophilized at a concentration of 1 mg / ml (dissolved in 25mMTBS / 50% glycerol) and stored at -20°C. The following...
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