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Duck circovirus and adenovirus bivalent inactivated vaccine and preparation method of yolk antibody thereof

A dual inactivated vaccine and duck circovirus technology, which is applied in the field of poultry epidemic immunization control, can solve problems such as large market vacancies, achieve the effect of improving antibody titer, optimizing immunization procedures and extraction steps, and good protection effect

Inactive Publication Date: 2020-07-17
山东百瑞凯来生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the limitations of duck circovirus vaccine and adenovirus vaccine production technology, there is no commercialized vaccine on the market at present, and there is a huge market gap. The egg yolk antibody used to prevent and treat early infection has not been reported.

Method used

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  • Duck circovirus and adenovirus bivalent inactivated vaccine and preparation method of yolk antibody thereof
  • Duck circovirus and adenovirus bivalent inactivated vaccine and preparation method of yolk antibody thereof
  • Duck circovirus and adenovirus bivalent inactivated vaccine and preparation method of yolk antibody thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] The preparation of embodiment 1 vaccine

[0053] The original virus species of circovirus was isolated from the liver of ducks with clinical onset in 2019. PCR detection technology was used to confirm that the diseased ducks were infected with duck circovirus, and other pathogens were detected to rule out the possibility of virus mixed infection, and then homogenized and centrifuged. The supernatant was filtered and sterilized with a disposable filter; the virus seed infected 1-day-old ducklings, and was continuously passed down for 3 generations. After the virus amount was detected to be qualified, it was used as a purified virus seed. The inventor carried out biological preservation of the virus strain, and the preservation number: CGMCC No.19296;

[0054] Adenovirus (serotype IV) strains were purchased from China Veterinary Microbiology Culture Collection Center (preservation number: CVCCAV211);

[0055] The 1-day-old healthy Cherry Valley ducklings were selected to...

Embodiment 3

[0084] Embodiment 3 Vaccination dose and safety evaluation

[0085] According to the conclusion drawn in Example 2, according to duck circovirus and adenovirus antigen ratio 3:1, pine pollen polysaccharide concentration 20mg / mL, produce vaccine, and evaluate the vaccination dose and safety of vaccine. The vaccination dose was set as three doses of 0.2, 0.5, and 1.0 mL, and each dose was inoculated with 5 ducklings at the age of 1, 4, and 7 days (only once inoculated, no additional immunization). Venous blood was collected, serum was separated, and the titer of duck circovirus antibody was detected by agar diffusion test, and the result was expressed as the mean value ± SD of titer log2. The stress situation of each group after inoculation was recorded, and the stress manifested as depression, reluctance to move around, reduced food intake, etc., and death in severe cases.

[0086] The results in Table 3 show that the age of inoculation has little effect on antibody production...

Embodiment 4

[0092] The making of embodiment 4 yolk antibody

[0093] (1) Use the vaccine used in Example 3 to immunize the laying hens 1 month before the start of laying in 4 times, the inoculation dose is 1.5mL / only, and the interval between each immunization is 2 weeks, and the follow-up is every 1-2 months immune once;

[0094] (2) Collect immunized eggs two weeks after the fourth immunization, and keep immunizing once every 1-2 months during the continuous egg harvesting process; separate the egg yolk from the egg white with an egg yolk separator, and separate the egg yolk from the egg yolk preheated to 35°C. Dilute the egg yolk liquid with purified water at a volume ratio of 3.5:1, stir evenly to obtain the egg yolk dilution, and pre-warm at 35°C for 1 hour.

[0095] (3) Add PEG6000 with a final concentration of 3.5% to the egg yolk dilution, mix it with the egg yolk, stir well, and let it stand for 4 hours to fully react.

[0096] (4) Take out the supernatant after the reaction, a...

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Abstract

The invention relates to a duck circovirus and adenovirus bivalent inactivated vaccine and a preparation method of a yolk antibody thereof. A duck circovirus antigen is obtained by infecting 1-day-oldhealthy cherry valley ducklings and collecting infected duck livers at the age of 25 days to obtain antigen tissues, and an adenovirus antigen is obtained by infecting SPF chickens of 20 days old to30 days old and collecting livers of dead chickens; an antigen of the vaccine consists of liver tissues infected with the duck circovirus and liver tissues infected with the adenovirus in a mass ratioof (1-5):1, and an antigen solution for the vaccine is prepared; and a pine pollen polysaccharide with a concentration of 5-40mg / mL is added into the antigen solution to serve as an immunopotentiator, and the antigen solution is emulsified with a conventional white oil adjuvant to obtain the vaccine. The yolk antibody is obtained by immunizing laying hens with the vaccine, and then performing extracting and purifying on egg yolks of high-immunity laying hens, and the yolk antibody simultaneously contains two antibodies against the duck circovirus and the adenovirus. The vaccine and yolk antibody provided by the invention have the advantages of simple preparation process, low cost, good action effect, stable dosage form and easy storage, and has a wide application prospect.

Description

technical field [0001] The invention relates to the field of immune control of poultry epidemic diseases, in particular to a preparation method of duck circovirus and adenovirus dual inactivated vaccine and egg yolk antibody thereof. Background technique [0002] Duck circovirus (Duck circovirus, DuCV) is a new member of the Circoviridae genus Circovirus, with no envelope and a single-stranded circular DNA structure. The virus was first reported by Hanemann in Germany in 2003. In my country, Jiang Shijin et al. (2007) detected DuCV for the first time in samples of Riemerella anatipestifer diseased ducks collected in Fujian Province in 2006. Liu Shaoning et al. (2008-2009) tested 343 tissue samples from 36 duck flocks in 5 provinces of Shandong, Jiangsu, Sichuan, Fujian and Guangdong, and found that the detection rate of DuCV was as high as 81.63%, and the positive rate of duck flocks was 94.44%; The positive rate of 18 meat duck farms in Shandong was 88.89%, and the positi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12A61K39/235A61P31/20A61K39/39A61P37/04C07K16/08C07K16/02
CPCA61K39/12A61K39/39A61K2039/5252A61K2039/552A61K2039/55583A61K2039/70A61P31/20A61P37/04C07K16/02C07K16/081C12N2710/10234C12N2750/10034
Inventor 刘金龙丁树新庄晓峰滕军
Owner 山东百瑞凯来生物科技有限公司
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