| Streptococcus suis is an important zoonotic pathogen, causing arthritis, meningitis,septicemia, bronchitis in pigs and pleurisy, septicemia, even death in human’s, not only caused great economic losses to aquaculture but also caused great concern in public health. Streptococcus suis can be divided into 35 serotypes according to the differences in the capsular polysaccharide antigen,serotype 1.1/2,2,7,9,14 are the main pathogenic serotypes among them.What’s more,the type 2 is the most popular and pathogenic which is used as a representative of the research of streptococcus pneumoniae strains.In order to identify the pathogenic bacterium which leads the pigs of a hoggery of Hubei Province to acute death and control the disease, PCR identification,drug sensitivity test and animal pathogenicity test were carried on. But the isolated bacterium is not Streptococcus suis serotype 2 which can’t be used as a representative of the research of streptococcus pneumoniae strains. So the standard Streptococcus suis type 2 strain was introduced from other laboratory, and the related research on the 1910 RR protein of the two-component regulatory system 1910 HK/RR was made.The research was made as follows:(1) Isolation and identification of Streptococcus suis and it’s resistance against antibioticsThree shares each of the hydropericardium and lungs were took as the path-logical material of disease.On these six shares the separation experiment, serological identification,drug sensitivity test and animal pathogenicity test were carried on. The result indit is other serotype strain, not serotype 1 strain or serotype 2 strain, not serotype 7 strain or serotype 9 strain.The Streptococcosis suis has the deter-minate pathogenicity Which can cause mice to death. The Streptococcosis suis is highly sentitive to Cephalosporin, Penicillin, Ampicillin, Chloroamphenicol,Rifam-picin, Vancomycin, Amoxicillin, Ofloxacin and Levofloxacin, or intermediate sensitive to Ciprofloxacin, Enrofloxacin,Paediatric Compound Sulfamethoxazole and Tetracycline,but resistance to Polymyxin B Sulfate.Sulfamethoxazole and Lincomycin. The results suggest that the pathogenic bacterium of the hoggery of Hubei Province is Streptococcus suis and Cephalosporin, Penicillin,Ampicillin can be used for the treatment of the disease.(2) Prokaryotic expression of Streptococcus suis type 21910 response regulator protein and preparation of polyclonal antibodyThis experiment was conducted to express and prepare polyclonal antibody of 1910RR protein. response regulator of Streptococcus suis type 2 1910rr,gene was amplified by PCR.and inserted into expression vector pET-30a, the recombinant plasmid pET30a-1910rr was successfully expressed in E. Coli BL21(DE3). The protein was purified with nickel column. eluted with different concentrations of imidazole to determine the optimal concentration of elution.Then it was dialyzed by TE buffer and concentrated with sugar in the dialysis-bag. The concentration of the protein was measured by spectrophotometer. The rabbit was given subcutaneous injection of the 1910RR protein one time per two weeks.totally four times. The third and fourth immunization was given doubling dosage.Good immunological activity of protein was confirmed by Western blot test.The polyclonal antibodies of rabbit anti 1910RR protein was preparated. The 1910RR protein was expressed in the supernatant hrough optimized the condition.The polyclonal antibodies of rabbit anti 1910RR protein was 1:4 through detection of single agar diffusion test.The pathogenic bacterium of the hoggery of Hubei Province was isolated and reference for clinical medication and control the disease was provided. And the preparation of 1910RR protein polyclonal antibody was successful and can be used for function research of 1910RR protein -gene interactions. |
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