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Bovine infectious rhinotracheitis virus and mycoplasma bovis dual inactivated vaccine and its preparation method and suspension mdbk cells used

A technology of rhinotracheitis virus and double inactivated vaccine, applied in the directions of virus/phage, virus antigen components, biochemical equipment and methods, etc., can solve the problem of affecting the effect of vaccine use, not involving the prevention of Mycoplasma bovis, and increasing animal stress and other problems, to achieve the effect of saving economic costs, high sensitivity, and ensuring stability

Active Publication Date: 2022-07-19
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Patent documents such as CN110420323A, CN109022314A, CN107050451A about the Mycoplasma bovis vaccine are all single vaccines at present, and patent documents such as CN106729692B about the IBRV vaccine all do not involve the prevention of Mycoplasma bovis, if want to prevent and control these two kinds of pathogens simultaneously need to immunize separately, this is both Increases the stress of animals and also affects the effectiveness of vaccines

Method used

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  • Bovine infectious rhinotracheitis virus and mycoplasma bovis dual inactivated vaccine and its preparation method and suspension mdbk cells used
  • Bovine infectious rhinotracheitis virus and mycoplasma bovis dual inactivated vaccine and its preparation method and suspension mdbk cells used
  • Bovine infectious rhinotracheitis virus and mycoplasma bovis dual inactivated vaccine and its preparation method and suspension mdbk cells used

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078]Example 1 Acquisition of MDBK cells capable of suspension culture of bovine infectious rhinotracheitis virus

[0079] 1. Screening of serum-free suspension medium

[0080] Set up 5 treatments in 5 suspension media:

[0081] Treatment A: Suspension medium was prepared by using the ST cell suspension medium of Eternal Life Sciences.

[0082] Treatment B: The suspension medium was prepared by using the Universal Suspension Cell Culture Medium of Yishengke.

[0083] Treatment C: Suspension medium was prepared using MDBK cell serum-free medium.

[0084] Treatment D: Suspension medium was prepared using CD MDBK 249.

[0085] Treatment E: Suspension medium was configured with OPM-MDBK SFM1 DPM.

[0086] For each treatment, the above serum-free medium and DMEM basal medium were mixed at a volume ratio of 1:1, and FBS and PS (penicillin-streptomycin mixture) were added to obtain a medium called suspension medium. The volume percentage of FBS in the base is 3%, the content of...

Embodiment 2

[0116] Example 2 The acquisition of the double inactivated vaccine of bovine infectious rhinotracheitis virus and mycoplasma bovis

[0117] 1. Preparation of IBRV antigen in suspension culture

[0118] 1.1 Preparation of vaccine strains

[0119] Suspended MDBK cells from Example 1 at an initial density of 1 × 10 6cells / ml, cultured in 125ml shake flasks (25ml per flask), after culturing for 48h, centrifuged at 300g (or 1000rpm) for 10min, using fresh serum-free suspension medium (preheated at 37°C for 30min) in 2.2.1 of Example 1 ) to dilute the cells to 4×10 6 cells / ml (25-30ml per flask), placed in a new cell shaker flask. The IBRV-SZH strain was inoculated into the shake flask of suspension cell culture at MOI=0.1 to carry out adaptive culture, and the shake flask was placed in 5% CO. 2 3. Culture on a shaker in a constant temperature incubator at 37°C, adjust the speed of the shaker to 100 rpm, and continuously cultivate for three generations, each generation for 48-72...

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Abstract

The present invention provides a double inactivated vaccine of bovine infectious rhinotracheitis virus and mycoplasma bovis, a preparation method thereof, and suspended MDBK cells used therein. The invention provides suspension MDBK cells, and on this basis, establishes a method for suspending IBRV antigen virus liquid. The IBRV antigen virus liquid obtained by the method has the characteristics of high antigen content, large antigen batch, stable batch, etc. The side reactions caused by the bovine serum added to the culture medium can save the economical cost while producing a safer and more efficient vaccine. At the same time, the IBRV and MB double inactivated vaccine of the present invention can prevent two diseases with one injection, simplify the immunization procedure, reduce the number of immunizations of animals, reduce the stress of animals, save labor, and further reduce the cost of animal breeding.

Description

technical field [0001] The invention relates to a double inactivated vaccine of bovine infectious rhinotracheitis virus and mycoplasma bovis in the technical field of veterinary biological products, a preparation method thereof, and suspended MDBK cells used therein. Background technique [0002] Bovine kidney cells (MDBK), a passaged cell line derived from apparently normal bovine kidneys, usually adherently grown in medium containing fetal bovine serum, are resistant to bovine infectious rhinotracheitis virus (IBRV). ), bovine viral diarrhea / mucosal virus (BVDV) and other viruses are more sensitive. [0003] Bovine infectious rhinotracheitis virus (IBRV), also known as bovine herpes virus type I (BHV-1), can cause symptoms such as high fever, loss of appetite, cough, runny nose and conjunctivitis in dairy and beef cattle, resulting in bovine infectious rhinotracheitis . [0004] Mycoplasma bovis (MB) can cause diseases related to the respiratory system of cattle, such as...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/295A61K39/02A61K39/265A61P31/04A61P31/22C12N5/071C12N5/02C12N7/00C12R1/93
CPCA61K39/0241A61K39/12A61P31/04A61P31/22C12N5/0686C12N7/00A61K2039/70A61K2039/5252A61K2039/521C12N2710/16734C12N2500/90C12N2500/84C12N2527/00C12N2710/16751
Inventor 吴文学王朋朋格勒图
Owner CHINA AGRI UNIV
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