Recombinant brucella expressing VP1 gene of O-type foot-and-mouth disease virus and method for producing vaccines thereof
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A technology of foot-and-mouth disease virus and Brucella, which is applied in the field of preventive live vaccines, can solve the problems of effect defects and achieve the effect of stable genetic traits
Active Publication Date: 2011-02-23
CHINA INST OF VETERINARY DRUG CONTROL
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Inactivated vaccines have been used in our country for decades, and the incr
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Embodiment 1
[0072] Pancreatic Soy Broth (TSB, BD Company, USA) or other suitable culture medium is used as the culture medium for recombinant bacteria, and an appropriate amount of commonly used antifoaming agent is added according to the medium volume, and 1% to 2% of the medium volume is added after sterilization. For the recombinant Brucella rS2-JS strain seed liquid, gradually increase the ventilation at 36-37°C, and ferment and cultivate for 36 hours according to the conventional method. 1% to 2% of the base amount.
[0073] After the cultivation is completed, try to use trypan soy agar to test the purity according to the method stipulated in "Chinese Veterinary Pharmacopoeia", and it is pure.
[0074] At the same time, take samples and count live bacteria with TSA for reference during concentration.
[0075] Add 0.2% to 0.4% of the total amount of sodium carboxymethyl cellulose to the bacterial liquid that has passed the pure inspection to precipitate the bacterial cells, absorb th...
Embodiment 2
[0080] Product testing
[0081] The relevant inspection methods involved in the present invention are carried out according to the method of "Chinese Veterinary Pharmacopoeia"
[0082] (1) Physical properties The vaccine should be a loose mass that can dissolve quickly (within 1 min) after adding the diluent;
[0083] (2) Pure inspection The vaccine should be purely free of other bacteria;
[0084] (3) Inoculate 18-20 g Kunming mice with a vaccine containing 500 million CFU live bacteria, and all of them should be healthy and alive within 6 days;
[0085] (4) Counting of viable bacteria Count the viable bacteria of the vaccine with TSA, and the number of viable bacteria per head is 80 billion to 100 billion CFU.
[0086] (5) Safety inspection Dilute rS2-Mya strain live vaccine with normal saline to 1ml containing 10 billion live bacteria, subcutaneously inject 5 mice weighing 18-20g, 0.1ml each, and all should be healthy and alive within 6 days.
[0087] (6) Immunogenicity...
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Abstract
The invention relates to recombinant brucella expressing a VP1 gene of O-type foot-and-mouth disease virus (FMDV) and a method for producing vaccines thereof. A recombinant strain is steadily integrated into the VPI gene of an O-type FMDV Myanmar strain from a brucella S2 strain genome, simultaneously, the condition of forming an O strand in a smooth type brucella cell wall LPS structure is destructed to ensure that the recombinant strain is changed from a smooth type into a rough type, the safety of the strain is further improved but a good immune effect on brucellosis is still kept, wherein the recombinant strain is named a recombinant brucella rS2-Mya strain. The recombinant strain can express the VP1 protein of the O-type FMDV Mya strain and induce to generate corresponding antibodies, and has good fundamental immunization effect on an O-type FMD. When the strain is used for producing the vaccines, the existing condition that brucella vaccine immune animals are difficult to distinguish from wild strain infected animals is changed, simultaneously the cellular immunity of FMD vaccines is realized, and good vaccines for preventing and controlling the brucellosis and the FMD are provided.
Description
[0001] Technical field The present invention relates to a live vaccine for preventing two infectious diseases of brucellosis and foot-and-mouth disease of cattle, sheep and pigs, and belongs to the field of veterinary biological products. technical background [0002] Brucellosis (brucellosis) is a zoonotic disease characterized by abortion and fever caused by Brucella or Brucella, which seriously threatens the life and health of humans and various animals. . This disease not only has serious harm to the reproduction and production performance of animals, but more importantly, people are often difficult to cure after being infected with Brucella, thus causing serious public health problems. Elimination of brucellosis has therefore been one of the most important goals of public health programs in countries where Brucella is endemic. At present, the main method to eliminate the disease worldwide is the combination of culling and immunization. For China, where the occurrence of ...
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