A kind of attenuated strain of Streptococcus suis type 2 and its application
A technology of Streptococcus suis and strains, applied in bacteria, antibacterial drugs, veterinary vaccines, etc., can solve the problems of strong virulence and reverse mutation of strains, and achieve the effect of reducing the incidence rate and improving the survival rate.
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Embodiment 1
[0025] Isolation and identification of Streptococcus suis type 2 XN
[0026] The disease material to be separated—the tissue disease material was obtained by the applicant from the lung tissue of pigs slaughtered in a slaughterhouse in Xianning, Hubei Province in November 2009. The specific separation method is as follows: aseptically collect the lung tissue and inoculate it on a TSA plate (purchased from BD Company, USA), and observe after culturing at 37°C for 12-24 hours. Pick small colonies with a diameter of 0.1mm-1.0mm, off-white, translucent, smooth, round, and with neat edges, and inoculate them in TSB liquid medium for overnight culture on a shaker at 37°C. Purely cultured bacteria were subjected to Gram staining for microscopic examination. Then observed with an optical microscope, Gram staining was positive, and it was subcultured and purified.
[0027] The classification and identification of Streptococcus suis type 2 was identified by PCR method and slide agglut...
Embodiment 2
[0038] Virulence test of Streptococcus suis type 2 XN
[0039] 1.1 Infection test of Streptococcus suis type 2 XN (SS2-XN) on mice
[0040] The pathogenicity of Streptococcus suis type 2 XN was determined using 4-week-old Kunming mice. The reported known virulent strain LT strain (CCTCC NO: M2011282) was used as a reference. Each bacterial strain was divided into 3 dosage groups (3×10 9 CFU, 5×10 8 CFU, 5×107 CFU), each dose group was subcutaneously inoculated with 10 mice, each inoculated with 0.5ml of bacterial suspension. After 7 days of observation, the statistical results are shown in Table 1.
[0041] Table 1 Differences in virulence of Streptococcus suis type 2 strains
[0042]
[0043] *High pathogenicity of HV: lethal dose ≤5×10 8 CFU, MV moderate pathogenicity: lethal dose is ≤3×10 9 CFU, LV low pathogenicity: 3×10 9 CFU is not lethal.
[0044] The results in Table 1 show that with 3×10 9 The SS2-XN strain of CFU was inoculated to 10 Kunming mice, and it...
Embodiment 3
[0052] Application of Streptococcus suis type 2 XN in the preparation of Streptococcus suis type 2 vaccine:
[0053] 1.1 Preparation of gelatin sucrose stabilizer
[0054] The gelatin sucrose protectant is a solution containing 12% (g / ml) gelatin and 40% (g / ml) sucrose.
[0055] 1.1.1 Accurately weigh 1200g of imported gelatin granules (Sigma Company), add it into a 10L liquid dispensing bottle, then inject about 8L of water for injection (pH value should be 6.8-7.2), stir well under the water temperature of 65°C, and dissolve evenly.
[0056] 1.1.2 Accurately weigh 4000g of sucrose (Sinopharm Group), add it to the above-mentioned liquid preparation bottle, and dissolve evenly under the same conditions.
[0057] 1.1.3 Make up the volume of the solution in the dosing bottle to 10L with water for injection.
[0058] 1.1.4 Sterilization Autoclave at 116°C for 30 minutes, set aside.
[0059] 1.1.5 Storage Store in a refrigerator at 2-8°C. Preheat in a 37°C incubator before use...
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