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Oral vaccine against clonorchiasis in fish and its preparation and application

A clonorchiasis sinensis and oral vaccine technology, which is applied to the oral vaccine of clonorchiasis sinensis and the fields of preparation and application thereof

Active Publication Date: 2021-05-07
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] At present, there are no patents and journal documents on the prevention of fish clonorchiasis vaccine at home and abroad

Method used

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  • Oral vaccine against clonorchiasis in fish and its preparation and application
  • Oral vaccine against clonorchiasis in fish and its preparation and application
  • Oral vaccine against clonorchiasis in fish and its preparation and application

Examples

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Embodiment 1

[0059] 1. Main solution preparation

[0060] Bacillus subtilis electroporation A medium: LB medium + 0.5M sorbitol, pH = 7.2.

[0061] Bacillus subtilis electroporation B medium: 0.5M sorbitol, 0.5M mannitol, 10% glycerol.

[0062] Bacillus subtilis electroporation C medium: 0.5M sorbitol, 0.5M mannitol.

[0063] LB liquid medium: NaCl 10g, yeast extract 5g, tryptone 10g, add ultrapure water to 1,000ml, adjust pH to 7.0, autoclave at 121°C for 20min.

[0064] LB solid medium: NaCl 10g, yeast extract 5g, tryptone 10g, agar powder 15g, add ultrapure water to 1,000ml, adjust pH to 7.0, autoclave at 121°C for 20min; wait until the temperature drops to about 50-55°C (Add the corresponding antibiotic solution when necessary), and pour the plate.

[0065] DSM Spore Medium (1,000ml): 8g Difco Nutrient Broth, 1g KCl, 0.25g MgSO 4 .7H 2 O, 0.002gMnCl 2 .4H 2 O; adjust pH=7.0, autoclave, add filter-sterilized 0.1M CaCl2 to a final concentration of 5×10 - 4 M; while adding filter...

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Abstract

The invention belongs to the field of biotechnology, and in particular relates to an oral vaccine against clonorchiasis sinensis and its preparation and application. The present invention firstly provides a fusion protein, containing Bacillus subtilis spore capsid protein CotC and Clonorchis sinensis cysteine ​​protease Cs CP. The fusion protein uses the Bacillus subtilis spore as a carrier to prepare an oral vaccine against clonorchiasis sinensis in adult fish, which can be successfully colonized in the intestinal tract of fish and has a good immune effect. The transmission route of clonorchiasis can be cut off, and the purpose of effectively preventing and controlling the prevalence and transmission of the disease can be achieved. Selecting Bacillus subtilis as the protein antigen presentation system can well avoid the defect that the pure protein is easily degraded by direct oral administration in the gastric acid environment of fish, and ensures that the vaccine can exert an effective protective effect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an oral vaccine against clonorchiasis sinensis and its preparation and application. Background technique [0002] Clonorchiasis, also known as liver fluke, is a foodborne zoonotic parasitic disease with the widest prevalence, the most threatened population, and the greatest harm in my country. Orchis sinensis (Clonorchis sinensis, C. sinensis) caused by freshwater fish or shrimp with metacercariae. Clonorchis sinensis infection can cause biliary tract inflammation, bile duct obstruction, and even cholangiocarcinoma, liver cirrhosis and liver cancer and other diseases. According to WHO reports, the risk of Clonorchis sinensis infection is 58 times higher than that of normal people, the risk of liver cirrhosis is 30 times higher than normal people, and the risk of liver cancer is 15 times higher than normal people. In March 2009, Clonorchis sinensis was identified by WHO a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/75A61K39/00A61P33/10
CPCA61K39/0003A61K2039/542A61K2039/55594C07K14/32C07K2319/00C12N9/6405C12N15/75C12N2800/101
Inventor 余新炳唐泽丽孙恒昌姜红烨黄艳陈庭金林志鹏周心怡
Owner SUN YAT SEN UNIV
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