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Canine distemper virus attenuated vaccine strain and application thereof

The technology of canine distemper virus and attenuated vaccine is applied in the field of prevention and treatment of canine distemper virus, which can solve problems such as no specific treatment, and achieve the effects of long immunization duration, good effect and early immunization generation period.

Active Publication Date: 2019-05-10
兆丰华生物科技(南京)有限公司 +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there is no specific treatment for this disease. The most effective measure to prevent and control this disease is regular vaccination of canine distemper vaccine.

Method used

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  • Canine distemper virus attenuated vaccine strain and application thereof
  • Canine distemper virus attenuated vaccine strain and application thereof
  • Canine distemper virus attenuated vaccine strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Isolation and identification of canine distemper virus virulent strain HBv strain

[0024] The mink used in the experiment were purchased from healthy mink negative for canine distemper virus antibody.

[0025] Isolation and culture of virus in a mink farm in Hebei province, when the tissue supernatant prepared from the spleen and lymph nodes of mink suspected of developing canine distemper was inoculated into Vero cells for blind passage to the fourth passage, the cells showed confluent cytopathic changes characteristic of canine distemper virus, and the inoculated cells Granules in the cytoplasm were denatured and fused into syncytia.

[0026] In the immunofluorescence test, the fifth-generation virus fluid was immunostained with a monoclonal fluorescent antibody against canine distemper virus. The cell control well had no specific green fluorescence, and the isolate inoculation well showed diffuse specific green fluorescence, which was positive. See figur...

Embodiment 2

[0035] Embodiment 2 passage of canine distemper virus weakens

[0036] Continuous passage of canine distemper virus Inoculate the monolayer of Vero cells with HBv strain of canine distemper virus, place at 37°C, 5% CO 2 Cultured under conditions, after confluent cytopathic lesions reached about 80%, the virus liquid was harvested by freezing and thawing, and recorded as the F1 generation. Such continuous passage to the F100 generation.

[0037] Determination of virus content The F20, F40, F60, F80, and F100 generation virus solutions were used for virus content determination, and the results are shown in Table 3.

[0038] Table 3 Virus content of different generations of canine distemper virus HBv strains

[0039]

[0040] Pathogenicity test on mink The F20, F40, F60, F80, F100 generation virus liquid was inoculated into 5 2-month-old mink respectively, and the 60th generation had no pathogenicity to the mink. The results are shown in Table 4.

[0041] Table 4 Pathogenic...

Embodiment 3

[0049] The preparation of embodiment 3 canine distemper live vaccine

[0050] Preparation of Canine Distemper Virus Antigen Canine distemper virus seed was inoculated into Vero cell monolayer at a ratio of 2%, cultured at 33°C, and when the cytopathic changes reached more than 80%, the virus liquid was harvested by freezing and thawing, and the virus content reached 10 4.5 TCID 50 / ml, it can be used for the preparation of canine distemper live vaccine after passing the purity test.

[0051] Preparation and inspection of canine distemper live vaccine Mix canine distemper virus antigen and suitable lyoprotectant (sucrose gelatin lyoprotectant) according to the volume ratio of 5:1, then quantitatively dispense vaccine bottles and lyophilize, virus content up to 10 4.0 TCID 50 / head portion, the remaining moisture, vacuum degree, sterility, mycoplasma, and exogenous virus inspection are qualified vaccines.

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Abstract

The invention discloses a canine distemper virus attenuated vaccine strain and application thereof. The canine distemper virus is named as an HBa strain, and the microorganism has the preservation number of CGMCC No.15036. The separated canine distemper virus is subjected to passage and cloning on a Vero cell, the attenuated vaccine strain (HBa strain) is cultured, and the results of a pathogenicity test and an immunogenicity test show that the canine distemper virus HBa strain is not pathogenic to both minks or dogs and can provide quite good protecting force for other sources of strong poison attacks. The attenuated vaccine strain has good immunogenicity, can be prepared into a single vaccine or combined vaccine, can effectively prevent diseases caused by the canine distemper virus, andcan also be used for preparing reagents or kits for treating, diagnosing or detecting the diseases caused by the canine distemper virus. The attenuated vaccine strain has the advantages of being stable in hereditary, lasting in immune, good in effect, safe, reliable, long in storage life and the life.

Description

technical field [0001] The present invention relates to an attenuated vaccine strain HBa strain which is attenuated by the virulent strain of canine distemper virus (CDV), and also relates to the preparation of the attenuated vaccine strain for preventing diseases caused by canine distemper virus. The use in medicine and the use of the attenuated vaccine strain in preparing, treating, diagnosing or detecting diseases caused by canine distemper virus belong to the field of prevention and treatment of canine distemper virus. Background technique [0002] Canine distemper virus is the pathogen that causes canine distemper in canines, mustelidae and some raccoons or other carnivorous animals. After young unimmunized animals are infected with canine distemper virus, they often show an acute fatal course. Clinically, it is characterized by biphasic fever, conjunctivitis, severe respiratory and digestive system inflammation, and neurological symptoms in the later stage. Adult anima...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K39/175A61K39/23A61K39/155A61K39/205A61K39/235A61K39/215A61K39/02A61K39/295A61P31/14A61P31/20A61P31/04C07K16/10C07K16/06A61K39/42G01N33/569C12R1/93
Inventor 廖丽晶王海燕陈翠云满坤
Owner 兆丰华生物科技(南京)有限公司
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