Streptococcus suis disease-haemophilus parasuis disease-porcine infectious pleuropneumonia triple subunit vaccine and preparation method thereof
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A technology for Haemophilus suis disease and Haemophilus suis, which is applied in the field of triple subunit vaccine and preparation thereof
Active Publication Date: 2021-09-07
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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At present, there is no triple subunit vaccine against Streptococcus suis, Haemophilus parasuis and porcine infectious pleuropneumonia in the world. Therefore, the present invention has strong innovation and application value, and has broad application prospects
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Embodiment 1
[0041] Expression and purification of embodiment 1 protein
[0042] 1 Extraction of bacterial genome
[0043] Streptococcus suis 05ZYH33 glycerol bacteria preserved in the laboratory were streaked in THA medium (containing 10% horse serum), then placed in a constant temperature incubator at 37°C overnight, and a single colony was picked and transferred to THB medium (containing 10% horse serum). % horse serum) in an incubator for static culture, and then expand the culture to 10mL THB medium (containing 10% horse serum), and use the bacterial genome extraction kit to extract the bacterial genome according to the instructions. The extracted genome was stored in a -20 refrigerator for future use.
[0044] Streak the Haemophilus parasuis HN10 glycerol bacteria preserved in the laboratory in TSA medium (containing 10 μg / ml NAD and 10% horse serum), then place it in a constant temperature incubator at 37°C overnight, and pick a single colony for transfer Put it into TSB medium (c...
Embodiment 2
[0066] Example 2 Preparation and Animal Test of Streptococcus Suis-Haemophilus Parasuis-Porcine Infectious Pleuropneumonia Triple Subunit Vaccine
[0067] 1 Preparation of triple subunit vaccine
[0068] Take the purified protein rMRP, rSLY, rEF, rCdtB, rAfuA, rOppA, rOppA2, rApxI, rApxII, rOMP mixed in Example 1, mix the mixed protein solution with ISA201VG in an equal mass ratio and emulsify, so that each dose of emulsified The content of each protein in the vaccine is 20 μg.
[0069] 2 Mouse Immunization Test
[0070] Twenty 4-week-old SPF C57 mice were randomly divided into 2 groups, 10 in each group, and 40 4-week-old SPF BALB / c mice were randomly divided into 2 groups, 20 in each group. Mice were immunized according to the immunization schedule in Table 5. The second immunization was carried out 14 days after the first immunization, and the dose and route of immunization were the same as those of the first immunization. 14 days after the second immunization, blood wa...
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Abstract
The invention discloses a streptococcus suis disease-haemophilus parasuis disease-porcine infectious pleuropneumonia triple subunit vaccine and a preparation method thereof. The triple subunit vaccine contains streptococcus suis antigen proteins MRP, SLY and EF, haemophilus parasuis antigen proteins AfuA, OppA2, CdtB and OppA, and actinobacillus pleuropneumoniae antigen proteins ApxI, ApxII and OMP. Experiments prove that the vaccine can stimulate strong immune response of mice, and has a good cross protection effect on streptococcus suis, haemophilus parasuis and actinobacillus pleuropneumoniae challenge mice with different serotypes, which fully shows that the triple subunit vaccine prepared in the invention has a good protection effect. Therefore, the invention lays a solid foundation for developing an efficient, broad-spectrum and cheap streptococcus suis disease-haemophilus parasuis disease-porcine infectious pleuropneumonia vaccine, and provides an effective technical means for preventing and treating streptococcus suis disease, haemophilus parasuis disease and porcine infectious pleuropneumonia.
Description
technical field [0001] The invention relates to a triple subunit vaccine and a preparation method thereof, in particular to a streptococcosis suis-Haemophilus parasuis-porcine infectious pleuropneumonia triple subunit vaccine and a preparation method thereof. The invention belongs to the technical field of veterinary medicine. Background technique [0002] Streptococcus suis is a bacterial infectious disease caused by Streptococcus suis (SS), which is characterized by clinical symptoms such as meningitis, septicemia, and arthritis in pigs. It has been a major problem that has plagued the global pig industry for a long time . There are many serotypes of Streptococcus suis, among which Streptococcus suis serotype 2 (SS2) is also a zoonotic pathogen. Humans can be infected with the pathogen through specific transmission routes (such as wound exposure, etc.) If it is not timely, it will cause sepsis, meningitis, etc., causing poor prognosis and seriously threatening public hea...
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