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BVDV (Bovine Virus Diarrhea Virus) SMU-Z6/1a/SC/2016 isolate and application thereof

A technology of bovine viral diarrhea and SMU-Z6, which is applied in the field of bioengineering to achieve the effects of good immunogenicity, strong pertinence and broad market application prospects

Active Publication Date: 2018-10-26
SOUTHWEST UNIVERSITY FOR NATIONALITIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, there are few commercial BVDV vaccines put into use in my country, and more importantly, the genotypes and subtypes of BVDV strains in my country are gradually increasing.

Method used

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  • BVDV (Bovine Virus Diarrhea Virus) SMU-Z6/1a/SC/2016 isolate and application thereof
  • BVDV (Bovine Virus Diarrhea Virus) SMU-Z6/1a/SC/2016 isolate and application thereof
  • BVDV (Bovine Virus Diarrhea Virus) SMU-Z6/1a/SC/2016 isolate and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The isolation and identification of embodiment 1 bovine viral diarrhea virus

[0032] 1.1 Isolation of bovine viral diarrhea virus

[0033] First observe the morbidity of the cattle herd, and the inventor collects a cattle feces sample initially diagnosed as diarrhea. Stool samples were diluted 1:5 with PBS (100 U / mL penicillin, 100 g / mL streptomycin). Centrifuge at 3000r / min for 10min, and the obtained supernatant is filtered and sterilized by a 0.22μm filter membrane to obtain a disease material treatment solution.

[0034] Inoculate the above-mentioned disease material treatment solution on a vigorously growing monolayer of MDBK cells on a 6-well culture plate, inoculate 0.5 mL per well, and set up negative control wells at 37°C with 5% CO 2 Incubate for 1 hour in the incubator, discard the inductive solution, add 2 mL of maintenance solution (DMEM with 1% fetal bovine serum), at 37°C containing 5% CO 2 Cultivate in an incubator, observe the cytopathic changes eve...

Embodiment 2B

[0052] The development of embodiment 2BVDV inactivated vaccine

[0053] 2.1 Virus reproduction

[0054] Take the MDBK monolayer cells in a good growth state, wash them twice with Hanks, inoculate the cell culture medium of the above-mentioned vaccine candidate strain SMU-Z6 / 1a / SC / 2016 with good immunogenicity into the MDBK cells, add 1% fetal bovine Serum in DMEM cell maintenance solution at 37°C with 5% CO 2 Cultivate in an incubator, and when the cytopathic rate is about 80%, harvest the virus culture medium, freeze and thaw 3 times, and centrifuge at 4°C at high speed to obtain the virus supernatant as the virus for the vaccine.

[0055] 2.2 Preparation of virus inactivated vaccine

[0056] The cell culture supernatant of the vaccine candidate strain SMU-Z6 / 1a / SC / 2016 with good immunogenicity above was fully mixed with β-propiolactone with a final concentration of 0.5%, and shaken at 37°C. Bed, inactivation treatment 6h. Two copies of the inactivated virus solution were...

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Abstract

The invention discloses a BVDV (Bovine Virus Diarrhea Virus) SMU-Z6 / 1a / SC / 2016 isolate and application thereof. A preservation number of BVDV is CCTCC (China Center for Type Culture Collection) NO: V201813, and the BVDV SMU-Z6 / 1a / SC / 2016 isolate is preserved in CCTCC. The BVDV has higher virus titer, a BVDV strain has good immunogenicity, an inactivated vaccine can be prepared from inactivated BVDV and an 201 adjuvant and is capable of immunizing calf and is capable of generating a higher antibody, and the BVDV strain is a good BVDV vaccine candidate strain. A diagnostic reagent, the inactivated vaccine and an attenuated vaccine which are relevant to the BVDV can be prepared on the basis of the BVDV, and the BVDV SMU-Z6 / 1a / SC / 2016 isolate has a wide market application prospect in diagnosison BVDV regional epidemic diseases and vaccine prevention and control.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a bovine viral diarrhea virus SMU-Z6 / 1a / SC / 2016 isolate and application thereof. Background technique [0002] Bovine virus diarrhea virus (BVDV) is one of the important pathogens that cause diarrhea in cattle. Infection of cattle with BVDV can lead to decreased milk production, reduced fecundity, growth retardation and secondary infection of other diseases. An acute, febrile, contact infectious disease characterized by diarrhea and mucous membrane necrosis, erosion or ulceration of the digestive tract. BVDV infection in cattle can manifest as clinical types such as subclinical infection, acute infection, mucosal disease and persistent infection. The World Organization for Animal Health (OIE) has classified bovine viral diarrhea mucosal disease as a Class B infectious disease, which has caused immeasurable economic losses to the world's cattle industry. Due t...

Claims

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Application Information

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IPC IPC(8): C12N7/00C12N7/06C12N7/02C12Q1/70C12Q1/686G01N33/569A61K39/12A61P31/14C12R1/93
CPCA61K39/12A61K2039/5252A61K2039/5254A61K2039/552A61P31/14C12N7/00C12N2770/24321C12N2770/24334C12N2770/24351C12N2770/24363C12Q1/686C12Q1/701G01N33/56983
Inventor 张斌阮文强汤承岳华王远微张丹丹
Owner SOUTHWEST UNIVERSITY FOR NATIONALITIES
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