Multivalence recombinant protein vaccine for chicken coccidiosis, and preparation method and application of multivalence recombinant protein vaccine for chicken coccidian

A technology of recombinant protein and chicken coccidia, which is applied in multivalent vaccines, immunoglobulins, immunoglobulins from serum, etc., can solve the problems of difficult popularization and application and high production cost

Inactive Publication Date: 2020-08-11
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the production cost is very high and it is difficult to popularize and apply

Method used

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  • Multivalence recombinant protein vaccine for chicken coccidiosis, and preparation method and application of multivalence recombinant protein vaccine for chicken coccidian
  • Multivalence recombinant protein vaccine for chicken coccidiosis, and preparation method and application of multivalence recombinant protein vaccine for chicken coccidian
  • Multivalence recombinant protein vaccine for chicken coccidiosis, and preparation method and application of multivalence recombinant protein vaccine for chicken coccidian

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Expression and purification of embodiment 1 recombinant protein

[0045] In the present invention, the antigen of Eimeria tenella is EtROP. The accession number in the ToxoDB database is ETH_00005405 (https: / / toxodb.org / toxo / app / record / gene / ETH_00005405), the length of the gene fragment encoding the protein The accession number of EtGAM22 in the ToxoDB database is ETH_00035480 (https: / / toxodb.org / toxo / app / record / gene / ETH_00035480), and the length of the gene fragment encoding the protein is 597bp; the accession number of EmGAM56 in the ToxoDB database The number is EMWEY_00026710 (https: / / toxodb.org / toxo / app / record / gene / EMWEY_00026710), and the length of the gene fragment encoding the protein is 1422bp; the accession number of Ea3-1E in the ToxoDB database is EAH_00057690 (https: / / toxodb.org / toxo / app / record / gene / EAH_00057690), the length of the gene fragment encoding the protein is 1737bp.

[0046] 1. Cloning of genes and construction of recombinant plasmids

[0047]...

Embodiment 3

[0085] Example 3 Evaluation of Maternal Immunization on the Protective Effect of Seven-day-old Offspring Chicks

[0086] 1. Preparation of sporulated oocysts of Eimeria gallinarum

[0087] Five 14-day-old coccidian-free chickens in each of the three groups were orally infected with 1×10 4 sporulated E.tenella sporulated oocysts, 2×10 5 sporulated oocysts of E.maxima and 1.5×10 5 Each group of sporulated oocysts of E.acervulina was reared in different isolators. After 144h, 121h and 97h, the feces of each group were collected for 3 consecutive days, and the oocysts were collected by floating in saturated saline (method omitted). The collected oocysts were cultured on a shaker at 27°C for sporulation. During this period, an appropriate amount of oocyst liquid was taken to examine the degree of sporulation under a microscope. When 95% of the oocysts were completely sporulated, they were stored at 4°C for later use.

[0088] 2. Evaluation of the protective effect of 7-day-old ...

Embodiment 4

[0148] Example 4 Evaluation of the protective effect of maternal immunization on 14-day-old offspring chicks

[0149] 1. The preparation method of fresh Eimeria chicken fresh oocyst: with embodiment 3;

[0150] 2. Evaluation of the protection effect of 14-day-old offspring chicks

[0151] 2.1 Experimental design

[0152] Eggs from the hen after three vaccinations were selected for incubation, and the eggs of the non-immunized hen were used for subsequent incubation in the challenge control group and the blank control group. The experiment was divided into 4 groups, with 10 14-day-old chicks in each group, randomly divided into groups, and the chicks with low or high body weight were eliminated. The first group of maternal immunization group (10 feathers), the second group of adjuvant immunization group (10 feathers), the third group of challenge control group (positive control group) (10 feathers), the fourth group of blank control group (negative control group). At the ag...

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Abstract

The invention discloses a multivalence recombinant protein vaccine for chicken coccidiosis, and a preparation method and application of the multivalence recombinant protein vaccine for chicken coccidian. The multivalence recombinant protein vaccine for chicken coccidiosis comprises 4 chicken coccidian proteins: an eimeria tenella rhoptry protein 41, an eimeria tenella gametophyte protein 22, an eimeria maxima gametophyte protein 56 and an eimeria acervulina merozoite protein Ea3-1E. The four chicken coccidian proteins are respectively connected with escherichia coli expression carriers to obtain recombinant expression carriers for expressing recombinant proteins. Different recombinant proteins are mixed with an adjuvant in proportion, when descendant chicks of immunized hens are inoculatedwith coccidian are compared with descendant chicks of hens which are not immunized, the survival rate and the relative weight increment of chicks inoculated with the coccidian can be effectively increased, the discharge quantity of oocysts is reduced, and favorable maternal immunity effects are shown.

Description

technical field [0001] The invention relates to the technical field of biological veterinary medicine, in particular to a chicken coccidia multivalent recombinant protein vaccine and its preparation method and application. Background technique [0002] Chicken coccidiosis (Coccidiosis) is one of the diseases that seriously endanger the chicken industry caused by seven kinds of Eimeria coccidiosis, and it is distributed worldwide. The pathogens causing chicken coccidiosis include Eimeria tenella, Eimeria maxima, Eimeria acervulina, Eimeria brucei brunetti), Eimeria necatrix, Eimeria praecox, and Eimeria mitis. Different species of coccidia parasitize different intestinal epithelial cells of chickens, causing a series of hazards such as enteritis, hemorrhage, decreased weight gain and even death. In production, chicken coccidiosis is often a mixed infection, the incidence rate is as high as 50-70%, the mortality rate is 20-30%, and when it is severe, it is as high as 80%. A...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/012A61P33/02C12N15/11C12N15/66C12N15/70C07K16/20C07K16/06
CPCA61K39/012A61P33/02C12N15/66C12N15/70C07K16/20C07K16/06A61K2039/70A61K2039/552
Inventor 刘群杨旭陈庆忠宋星桔刘晶安同伟许建海
Owner CHINA AGRI UNIV
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