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A kind of preparation method of goose gout yolk antibody

A yolk antibody, goose gout technology, applied in biochemical equipment and methods, virus/phage, antiviral immunoglobulin, etc. The effect of enhancing the immune function of the body

Active Publication Date: 2022-02-25
TIANJIN RINGPU BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Another object of the present invention is to provide a yolk antibody for preventing and treating goose gout, so as to solve the problem that there is no medicine available for goose gout in the existing clinics.

Method used

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  • A kind of preparation method of goose gout yolk antibody
  • A kind of preparation method of goose gout yolk antibody
  • A kind of preparation method of goose gout yolk antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Isolation and Identification of Wild Goose Astrovirus

[0045] Lesioned livers and kidneys from goslings from clinical infection were homogenized with antibiotics in viral transport medium and the homogenized tissue was filtered through a 0.45 micron syringe filter. 0.1ml of virus liquid homogenate was continuously cultured on the primary goose embryo kidney cells with 0.1-2% serum for 4 generations, and the appropriate virus serum culture concentration was found, about 0.5-1%. With the development of 70%-80% cytopathic effect, cell cultures were frozen and stored at -80°C.

[0046] (1) RNA extraction. Using the RNA / DNA virus extraction kit (Takara), virus RNA was extracted from clinically infected gosling lesion liver and primary goose embryonic kidney cell cultures.

[0047] (2) Primer design. Utilizing the published goose Astrovirus sequence and the conserved ORF1b sequence of Astrovirus, primers were designed with Primer5.0 software. The primer sequenc...

Embodiment 2

[0071] Example 2 Preparation of Goose Astrovirus Venom and Inactivated Vaccine

[0072] 1. Virus expansion: 100-fold dilution of RPVA1601 virus seeds, inoculation of 100 susceptible goose embryos at the age of 10 days with chorioallantoic membrane, 0.2ml per embryo, incubation at 36-37°C, and inspection of embryos twice a day. Goose embryos that died within 48 to 240 hours after inoculation were placed at 2 to 8°C for 4 to 12 hours, and allantoic fluid, amniotic fluid and embryo bodies were collected, and the heads and limbs were removed from the embryo bodies. Homogenize with embryo fluid, freeze and thaw three times, centrifuge at 8000r / min for 20min, take supernatant and mix it in a sterile container, store at 2-8°C. (See Table 1).

[0073] Table 1 Preparation of venom

[0074] Strain name Embryo variety Number of embryos inoculated (pieces) Embryo age Harvest venom (ml) RPVA1601 susceptible goose embryo 100 10 1480

[0075] 2. Concentrati...

Embodiment 3

[0086] Example 3 Preparation of Goose Astrovirus Egg Yolk Antibody

[0087] 1. Preparation of highly immune eggs: immunize laying hens with the goose astrovirus inactivated vaccine prepared above, and inject 2.0ml of inactivated vaccine subcutaneously into the neck of each chicken for the first immunization, and carry out the second immunization after 21 days. Inject 2.0ml of inactivated vaccine subcutaneously on the head, carry out the third immunization 21 days after the second immunization, inject 2.0ml inactivated vaccine subcutaneously in the neck of each chicken, and carry out booster immunization 21 days after the third immunization, and inject 2.0ml of inactivated vaccine subcutaneously in the neck of each chicken ml inactivated vaccine, 14 days after the booster immunization, the egg yolk was collected for challenge protection experiment.

[0088] 2. Production of egg yolk antibody:

[0089] (1) Eggshell disinfection: soak 18 kg of high-quality eggs in 0.1% bromogera...

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Abstract

A method for preparing goose gout yolk antibody. The present invention isolates goose astrovirus RPVA1061 strain from clinically infected goose diseased liver and kidney, uses RPVA1061 strain to prepare yolk antibody for the prevention and treatment of clinical goose gout, and has no drug use in solving the existing clinical goose gout disease. At the same time, a new clinical prevention and treatment scheme for goose gout is provided. Through clinical verification, the yolk antibody prepared by the present invention has a significant effect on the prevention and treatment of goose gout.

Description

technical field [0001] The invention belongs to the technical field of yolk antibody preparation, and in particular relates to a preparation method of yolk antibody for treating goose gout caused by astrovirus. Background technique [0002] From the beginning of 2017 to the present, an infectious disease with visceral and joint gout as the main symptoms has occurred in goose flocks in Shandong, Jiangsu, Guangdong, Guangxi, Hunan, Henan and Anhui, which has caused serious economic losses to the goose industry in my country. loss. The disease mainly occurs in goslings aged 1-20 days, and the mortality rate can reach up to 60%. Sick goslings are depressed, lying on the ground, tired and moving, and their food intake decreases. Necropsy showed severe urate deposition in internal organs and joint cavities, which occurred in different breeds, geese using different feeds, and different drugs. Reducing the protein content in the feed and reducing the amount of feed were ineffective...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/10C07K16/02C12N7/00
Inventor 李守军杨燚刘冠星郁宏伟刘海霞李蓬飞陈冰张立霞
Owner TIANJIN RINGPU BIO TECH
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