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Bovine viral diarrhea virus strain and application thereof

A bovine viral diarrhea and virus technology, applied in antiviral agents, virus/bacteriophage, antiviral immunoglobulin, etc., can solve the problem of pathogen screening and population purification in large-scale cattle farms, BVDV awareness and harm Insufficient attention is paid to the problem, and it is difficult to deal with clinical strains, etc., so as to achieve the effect of low cost, short production cycle, and easy fabrication

Active Publication Date: 2016-02-03
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the late start of research on BVDV in my country, insufficient attention has been paid to the awareness and hazards of BVDV, and no systematic nationwide molecular epidemiological survey of BVDV has been carried out. Research data; at the same time, BVDV is an RNA virus, which is prone to genetic mutations. It is difficult to prepare vaccines and detection kits with traditional standard strains to cope with all existing clinical strains; on the other hand, the BVDV detection kits imported from abroad are currently The price is expensive, and there is no self-developed commercial BVDV antigen detection kit in China, which makes it difficult to realize large-scale cattle herd pathogen screening and population purification on large-scale cattle farms

Method used

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  • Bovine viral diarrhea virus strain and application thereof
  • Bovine viral diarrhea virus strain and application thereof
  • Bovine viral diarrhea virus strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Virus isolation and identification.

[0025] 1. Clinical sample collection and BVDV detection

[0026] The inventor of the present invention investigated 25 cattle farms around Beijing from 2010 to 2013, collected and tested more than 4,000 clinical samples, and tested 1,000 randomly selected samples for antibody detection by indirect ELISA method. Antigen capture ELISA and RT-PCR methods were used to detect virus antigens, and 18 antigen-positive samples were detected. The detection method is as follows:

[0027] Reverse transcription reaction system: apply a two-step method for reverse transcription.

[0028] first step:

[0029] Random primer (250ng / μL) 1μL

[0030] vRNA (<500ng) 12μL

[0031] Mix well, centrifuge briefly at 65°C for 5min, and place on ice for 1min.

[0032] Step two:

[0033]

[0034]

[0035] Add the ingredients in the table in sequence, blow and mix gently, reaction conditions: 25°C for 5min, 50°C for 60min, 70°C for 15min. ...

Embodiment 2

[0048] The preparation of embodiment 2 immune antigen

[0049] 1. Seed poison preparation

[0050]Take the bovine kidney cell line MBDK cells preserved in the laboratory for culture, dilute the BVDV virus BJ1305 strain with DMEMF12 medium and inoculate monolayer cells, inoculate 1ml of each cell bottle, add cell culture medium, and place at 37°C, 5% CO 2 Cultivate in the cell culture box for 24 hours, collect the cell culture and freeze-thaw to obtain the breeding virus by ultrasonic crushing, the crushing power is 200W, 10min, and store the breeding virus in liquid nitrogen.

[0051] 2. Preparation of virus propagation

[0052] Culture MBDK cells in a single layer, inoculate the cell monolayer with a 100-fold dilution of the breeding virus, add complete cell culture solution containing 2% BVDV-free fetal bovine serum, and place at 37°C, 5% CO 2 Cultivate in the cell incubator for 24 hours, freeze and thaw several times, and mix the cells in each bottle. 3. Viral antigen co...

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Abstract

The invention relates to a bovine viral diarrhea virus BJ1305 strain, which is named as bovine viral diarrhea virus in a classifying manner, the preservation number is CGMCC NO.11185, the preservation date is September 7, 2015, and the preservation authority is General Microbiology Center of China Committee for Culture Collection of Microorganisms. The strain provided by the invention has the characteristics of being new in a region and epidemic, the immunogenicity is strong, and due to the fact that the train is separated from a Beijing area, products developed by utilizing the strain can specifically aim at prevalent diseases in the Beijing area, which is caused by the bovine viral diarrhea virus.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a bovine viral diarrhea virus strain and application thereof. Background technique [0002] Bovine Viral Diarrhea Virus (BVDV), also known as Bovine Viral Diarrhea-Mucosa Virus, is a serious disease that infects cattle with various clinical symptoms. Since Olafson et al first discovered the disease in New York State of the United States in 1946, it has been widely prevalent in the world and has caused serious economic losses to animal husbandry all over the world. Epidemiological survey results showed that BVDV was widely infected in Beijing, and persistent infection of cattle was the main cause of widespread infection. At the same time, the genetic differences of the epidemic strains were large, which brought great difficulties to the prevention and control of the disease. [0003] At present, the overall solution plan for the control and eradication of BVDV mainly includes compreh...

Claims

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Application Information

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IPC IPC(8): C12N7/00C07K16/10A61K39/395A61P31/14G01N33/569
Inventor 王九峰宋泉江翁晓刚才冬杰杜芊张旺
Owner CHINA AGRI UNIV
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