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Mink viral enteritis and canine distemper binary living vaccine as well as preparation method and application thereof

A technology for viral enteritis and canine distemper, applied in the field of veterinary biological products

Active Publication Date: 2015-05-06
华威特(江苏)生物制药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Domestic mink viral enteritis vaccines are all inactivated vaccines, and canine distemper is a live vaccine, and there is no combined vaccine

Method used

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  • Mink viral enteritis and canine distemper binary living vaccine as well as preparation method and application thereof
  • Mink viral enteritis and canine distemper binary living vaccine as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The source and characteristic of embodiment 1 system seedling poisonous seed

[0035] ① Mink viral enteritis virus JLM strain

[0036] The virus strain used to manufacture this product was isolated from mink feces swabs collected from a mink farm in Jilin Province. The strain was confirmed by CRFK cell passage, specific neutralization test, immunofluorescence test, PCR and gene sequence analysis. Mink viral enteritis virus (MEV). Animal regression tests showed that the strain had only weak pathogenicity. Then we subcultured the virus strain on CRFK cells to attenuate, and the virus titer could reach 10 5.6 TCID 50 / ml or more. The safety, immunogenicity, specificity, and purity of the MEV JLM strain virus were comprehensively identified, indicating that the MEV JLM strain virus has excellent safety, good immunogenicity, sterility, no mycoplasma, and no exogenous virus contamination.

[0037] The mink viral enteritis virus, named JLM strain, is classified as mink vi...

Embodiment 2 2

[0041] The preparation method of embodiment 2 double seedlings

[0042] 1. Preparation of virus liquid for seedling production

[0043] 1.1 Preparation of MEV virus liquid for seedling production

[0044] 1.1.1 Preparation by spinner bottle method

[0045] Well-grown CRFK cells were taken and simultaneously inoculated with MEV JLM virus (CGMCC No.9904) at an MOI of 0.005-0.5, and cultured at 37°C. After 4 to 5 days of cultivation, the virus liquid was harvested, frozen and thawed once at -15°C, and then stored. The virus content of each batch was measured respectively and sterility test was carried out for future use.

[0046] 1.1.2 Preparation by suspension culture method

[0047] Well-grown CRFK cells were inoculated into cell spinner bottles, and cultured at 37°C for 48-72 hours. The control speed is 8-12 rpm. When the cells grow into a dense monolayer, they are digested with 0.25% trypsin into single cells and counted, according to 0.8×10 5 ~1.2×10 5 Cell density pe...

Embodiment 3

[0067] Example 3 Mink Viral Enteritis, Canine Distemper Dual Live Vaccine Overdose Inoculation Safety Test

[0068] In order to evaluate the safety of mink viral enteritis and canine distemper dual live vaccine inoculation in mink, 20 healthy mink with negative viral enteritis, canine distemper antigen and antibody at the age of 7 to 8 weeks were used and divided into 4 groups. 5 in each group. Groups 1 to 3 are the vaccination groups, which were inoculated with 3 batches of the vaccine (batch numbers: 2008121, 2008122, and 2008123), 10 doses per animal (the normal dose is 1 dose per animal), and the fourth group was For the control group, each animal was inoculated with 1ml of PBS solution, and the temperature of the experimental animals was measured and clinically observed for 21 consecutive days after vaccination, and all animals were autopsyed at the end of the experiment. The results showed that after vaccination, the body temperature of the minks in each group was norma...

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Abstract

The invention discloses a mink viral enteritis and canine distemper binary living vaccine as well as a preparation method and application thereof. The effective components of the binary living vaccine comprise a mink viral enteritis virus (MEV) antigen and a canine distemper virus (CDV) antigen, wherein the MEV antigen is a JLM strain and has the collection number of CGMCC No. 9904, and the CDV antigen is a JTM strain and has the collection number of CGMCC No. 9905. Researches on the humoral immunity, the immunizing and virus attacking protection level and the pathologic change show that the JLM strain and the JTM strain are free from mutual immunity interference. The safety and effect of the binary living vaccine are researched further accordingly, and the research result shows that the binary living vaccine is safe to use and is not lower than the unitary living vaccine in effect.

Description

technical field [0001] The invention relates to a dual live vaccine and its preparation method and application, in particular to a mink viral enteritis and canine distemper dual live vaccine and its preparation method and application. The invention belongs to the technical field of veterinary biological products, Background technique [0002] As a precious fur animal, mink has been widely cultivated all over the world. Denmark, China, the United States, Canada, the Netherlands, Russia, and Finland are all major mink breeding countries, and my country's mink breeding volume ranks first in the world. Mink breeding has the characteristics of stable market, low operating risk and good economic benefits, and has a good breeding prospect. Since the mink breeding industry in my country is mostly small-scale family-style scattered farming, the breeding conditions are poor, the immune prevention and control is not uniform and not timely, and the epidemic of diseases has caused the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/295A61K39/175A61K39/23A61P31/20A61P31/14C12N7/00C12R1/93
Inventor 武华和彦良霍志云陈超阳
Owner 华威特(江苏)生物制药有限公司
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