Canine distemper and parvovirus disease bivalent inactivated vaccine for dogs and preparation method of canine distemper and parvovirus disease bivalent inactivated vaccine
A dual inactivated vaccine and canine distemper virus technology, applied in veterinary vaccines, biochemical equipment and methods, vaccines, etc., can solve the high infection rate and fatality rate of parvovirus in dog breeding and economic animal breeding losses, Immunization effects of live vaccines are greatly reduced and other issues
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Embodiment 1
[0032] The preparation of embodiment 1 canine distemper, parvovirus disease dual combination inactivated vaccine
[0033] (1) Inoculate the CDV-LG strain of canine distemper virus on Vero cells of African green monkey kidney cells to carry out virus amplification and culture, and harvest the virus liquid when the cytopathic changes reach 80%;
[0034] (2) Inoculate the canine parvovirus CPV-W strain on the cat kidney cell F81 to carry out virus amplification and culture, and harvest the virus liquid when the cytopathic changes reach 80%;
[0035] Specifically:
[0036] Preparation of poisonous seeds for production
[0037] Virus breeding
[0038] For the CDV-LG strain of canine distemper virus, dilute the virus seeds with MEM medium, absorb and inoculate Vero cells according to the MOI of 0.1-0.3 inoculum, cultivate at 37°C for 24 hours, change the maintenance medium, and cultivate at 33°C. % or more, freeze-thaw to collect the poison, and store below -20°C.
[0039] For t...
Embodiment 2
[0080] Example 2 Safety Test of Canine Distemper and Parvovirus Dual Inactivated Vaccine for Dogs
[0081] Get 3 batches of qualified canine distemper and parvovirus dual inactivated vaccines for dogs (prepared by the method of embodiment 1), inoculate healthy susceptible dogs, and establish single-dose group, single-dose repeated group and overdose (10 times) In the immunization group, dogs not vaccinated were set as controls. After vaccination, observe continuously for 14 days to observe whether there is any abnormality in the dog's body temperature, appetite, mental state, and feces. No clinical symptoms were shown, and no lesions were observed in the main organs of the inoculated Beagle dogs, and there was no obvious inflammatory reaction at the inoculation site. It shows that the canine distemper and parvovirus dual inactivated vaccine is safe for dogs.
Embodiment 3
[0082] Example 3 Efficacy Test of Canine Distemper and Parvovirus Dual Inactivated Vaccine for Dogs
[0083] 1. Method
[0084] (1) Get canine distemper virus neutralizing antibody, canine parvovirus hemagglutination inhibitory antibody negative canine distemper, parvovirus disease double inactivated vaccine 3 batches (prepared by the method of embodiment 1), inoculate antibody negative dog, One head per animal, and a control group inoculated with cell culture medium, blood was collected every 3 months after immunization, and serum was separated for detection of neutralizing antibodies and hemagglutination inhibitory antibodies.
[0085] (2) 3 months, 6 months and 9 months after the dog was vaccinated, the canine distemper and parvovirus virulent viruses were used for the test respectively for challenge inoculation, and the challenge dose was 100 LD for each virulent virus 50 / Only. Observe the clinical symptoms such as body temperature, appetite, feces, and mental state of ...
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