Method by utilizing rabbit-origin cells to produce swine fever live vaccine
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A swine fever live vaccine and cell technology, which is applied in biochemical equipment and methods, microorganisms, pharmaceutical formulations, etc. Ensuring safety and high viral content
Inactive Publication Date: 2012-01-25
南京创启生物科技有限公司
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[0003] Because the infection rate of viral diarrhea virus (BVDV) in my country's cattle herd is very high, almost 100%, so the production of swine fever vaccine with bovine testicular cells is easy to cause cell exogenous virus contamination, and the virus titer is not high. Unstable, large differences between vaccine batches, seriously affecting the quality of the vaccine; similarly, the use of sheep kidney cells to produce swine fever vaccine, due to the possible presence of border virus (BDV) in the flock, also has the same hidden danger
Bovine viral diarrhea virus (BVDV), sheep border virus (BDV) and swine fever virus (HCV) all belong to the genus Pestivirus in the family Flaviviridae. There is a close antigenic and serolog
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Embodiment 1
[0034] 1. Selection of cells for seedling production: the kidneys of healthy baby rabbits or adult rabbits;
[0035] 2. Preparation of cells for seedling production:
[0036] (1) Collection and processing of rabbit kidneys: first soak rabbit hair in normal water, drain it after taking it out, then soak it in 1% bromogeramine solution for 30 seconds, and after the body surface is sterilized, move it into a sterile room and use aseptic Rabbit kidneys were collected during the operation, quickly put into Hank’s solution containing 400IU / ml penicillin and 400μg / ml streptomycin, soaked for 30 minutes, and then removed the capsule, renal pelvis, and renal calices by aseptic operation, and weighed the tissues. Cut it into small pieces of 1-2 mm, wash it repeatedly with Hank's solution until the supernatant is clear;
[0037] (2) Digestion: Move the tissue block into a digestion bottle, add 0.25% trypsin solution 5-6 times the volume ratio of the rabbit kidney mass, plug the bottle t...
Embodiment 2
[0050] 1. Selection of cells for making seedlings: the testis of healthy baby rabbits or adult rabbits;
[0051] 2. Preparation of cells for seedling production:
[0052] (1) Collection and processing of rabbit testicles: first soak the rabbit hair in normal water, drain it after taking it out, and then soak it in 1% bromogeramine solution for 30 seconds. Rabbit testes were collected during the operation, quickly put into Hank’s solution containing 400IU / ml penicillin and 400μg / ml streptomycin, soaked for 30 minutes, and then removed the capsule and epididymis by aseptic operation, weighed the tissue, and cut it into For small pieces of 1-2 mm, rinse repeatedly with Hank's solution until the supernatant is clear;
[0053] (2) Digestion: Move the tissue block into a digestion bottle, add 0.25% trypsin solution with a mass-volume ratio of 6-8 times the rabbit testis tissue, plug the bottle tightly, and place it in a water bath at 37°C for 60 minutes for digestion. During the d...
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Abstract
The invention relates to a method by utilizing rabbit-origin cells to produce swine fever live vaccine, which comprises the following steps that: selecting cells for producing vaccine, preparing cells for producing vaccine, breeding cell virus strain, breeding venom for producing vaccine, preparing vaccine, split charging and freeze drying. The method is characterized in that: rabbit-origin susceptible cells are used for substituting bovine testicular cells or goat kidney cells to produce swine fever live vaccine, which can solve the pollution problem of external virus from cow and sheep, and by strictly controlling the raw materials and the culture conditions, the purity of the produced vaccine can be guaranteed, and the safety and the effect of the vaccine can be guaranteed; and the rabbit-origin susceptible cells to substitute the transferred cells to manufacture the swine fever live vaccine, so the potential hazard that the transferred cells can cause cancer can be avoided, and the safety of the vaccine is higher. The swine fever cell venom cultured through the method has high content of virus, the antigen tilter is 10 times or more of that of the present vaccine, the immune efficacy of the vaccine can be greatly improved, and the weaknesses of the present vaccine in the clinical use can be overcome.
Description
technical field [0001] The invention belongs to the technical field of veterinary biological products, in particular to a method for producing live swine fever vaccine with rabbit-derived cells. Background technique [0002] Classical swine fever virus (CSFV) is a serious animal virus, which causes huge economic losses to the pig industry every year. At present, attenuated live vaccine immunization is mainly used in the world to prevent and control swine fever, so as to control the prevalence of swine fever. The attenuated CSF vaccine strain (strain C) propagated in heterologous primary cells is currently the main attenuated vaccine in my country. C strain is a vaccine strain with good immune protection that was bred from rabbits in the 1950s in my country. Because the vaccine strain is stable, safe and protective, it is recognized as the best and most widely used vaccine strain in the world. Due to the needs of application, the vaccine production of CSFV in my country ha...
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