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Recombinant brucella for expressing Asia type-I foot and mouth disease virus VP1 genes 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 and can solve problems such as defects in immune effects
Active Publication Date: 2012-05-23
CHINA INST OF VETERINARY DRUG CONTROL
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Problems solved by technology
Inactivated vaccines have been used in our country for decades, and the increasingly serious situation of FMD epidemic has shown the defect of its immune effect
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Embodiment 1
[0073] 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. Recombinant Brucella rS2-JS strain seed bacterial liquid, gradually increase the ventilation at 36-37°C, and ferment and cultivate for 28-38 hours according to the conventional method. The amount is 1% to 2% of the total amount of the culture medium.
[0074] After the cultivation is completed, try to use trypan soy agar (TSA, BD Company, USA) to test the purity according to the method stipulated in the "Procedures", and it is pure.
[0075] At the same time, take samples and count live bacteria with TSA for reference during concentration.
[0076] Add 0.2% to 0.4% of the total amount of sodium carboxymethyl cellulose to the bacterial liquid that has passed the pure inspectio...
Embodiment 2
[0081] Product testing
[0082] The relevant inspection methods involved in the present invention are carried out according to the method of the "Regulations"
[0083] (1) Physical properties The vaccine should be a loose mass that can dissolve quickly (within 1 min) after adding the diluent;
[0084] (2) Pure inspection The vaccine should be purely free of other bacteria;
[0085] (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;
[0086] (4) Counting of viable bacteria Count the viable bacteria of the vaccine with TSA, and the number of viable bacteria per head is between 80 billion CFU and 100 billion CFU.
[0087] (5) Safety inspection Dilute the rS2-JS strain live vaccine with normal saline to 1ml containing 10 billion CFU of live bacteria, subcutaneously inject 5 mice with a weight of 18-20g, 0.1ml each, and all should be healthy and alive on the 6th day.
[0088] (6)...
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Abstract
The invention relates to recombinant brucella for expressing Asia type-I foot and mouth disease virus VP1 genes and a method for producing vaccines thereof. Asia type-I foot and mouth disease virus (FMDV) Jiangsu strain VP1 genes are integrated into a brucella S2 strain genome, and the forming condition of an O chain in a smooth brucella cell wall LPS structure is simultaneously damaged so that the recombinant strain is changed into a rough type from a smooth type, the safety of the strain is further improved, but good immune effect on the brucella is still kept; and the recombinant strain is named as a recombinant brucella rS2-JS strain. The strain can express the Asia I FMDV VP1 protein, induces the generation of corresponding antibodies and has good basic immune effect on the Asia I FMD. The strain for producing the vaccines can change the current situation that brucella vaccine immunized animals and wild strain infected animals are difficult to distinguish, realizes cellular immunity of FMD vaccines at the same time, and provides a good vaccine for prevention and control of brucella diseases and FMD.
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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