Recombined enterotoxin (namely, fimbriae composite polyvalent vaccine) for resisting diarrhea of piglets and preparation method for recombined enterotoxin (namely, fimbriae composite polyvalent vaccine)

A technology for piglet diarrhea and multivalent vaccines, applied in antibacterial drugs, pharmaceutical formulas, bacterial antigen components, etc., can solve the problems of geographical restrictions on vaccine use, single antigen, and low immune protection rate

Inactive Publication Date: 2013-04-03
DALIAN UNIV OF TECH
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Problems solved by technology

[0006] The production problem to be solved by the present invention is that the vaccine against piglet diarrhea in the existing breeding production contains a single antigen, the immune protection rate is low,

Method used

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  • Recombined enterotoxin (namely, fimbriae composite polyvalent vaccine) for resisting diarrhea of piglets and preparation method for recombined enterotoxin (namely, fimbriae composite polyvalent vaccine)
  • Recombined enterotoxin (namely, fimbriae composite polyvalent vaccine) for resisting diarrhea of piglets and preparation method for recombined enterotoxin (namely, fimbriae composite polyvalent vaccine)
  • Recombined enterotoxin (namely, fimbriae composite polyvalent vaccine) for resisting diarrhea of piglets and preparation method for recombined enterotoxin (namely, fimbriae composite polyvalent vaccine)

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Embodiment Construction

[0028] Specific embodiments of the present invention will be described in detail below in conjunction with technical solutions and accompanying drawings.

[0029] 1. Vaccine preparation

[0030] 1. Primary strain culture and strain identification of standard pili strains

[0031] The standard K88 pili, K99 pili and F41 pili strains were inoculated on the modified Minca medium, cultured overnight at 37°C, and blood agglutination experiments were carried out with the corresponding (K88, K99, F41) pili antiserum, and the agglutination reaction reached " ++++", the pure inspection is qualified. The bacterial solution was added to 50% glycerol and frozen in a -70°C refrigerator.

[0032] 2. Secondary strain culture and extraction and identification of antigenic protein

[0033] (1) Extraction of recombinant enterotoxin protein SLS

[0034] The trivalent enterotoxin recombinant bacterium pET30-SLS was inoculated in TSB medium and cultured with shaking at 37°C. Microplate reader...

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Abstract

The invention discloses recombined enterotoxin (namely, fimbriae composite polyvalent vaccine) for resisting diarrhea of piglets and a preparation method for the recombined enterotoxin (namely, the fimbriae composite polyvalent vaccine), and belongs to the technical field of animal husbandry and veterinary. The composite vaccine comprising an enterotoxin protein antigen and a various fimbriae antigen protein is prepared by the following steps of: uniformly emulsifying recombined trivalent enterotoxin SLS proteins expressed by gene engineering recombination and fimbriae antigen proteins of the conventional pathogenic bacteria K88, K99 and F41 in China with an adjuvant. The recombined enterotoxin (namely, fimbriae composite polyvalent vaccine) comprises two important pathogenic factors of an ETEC (enterotoxigenic e.coli) pathopoiesia loop, namely, enterotoxin and fimbriae; each process of diarrhea generated by the ETEC is protected; in the first step, the growth of the ETEC is stopped; in the second step, the enterotoxin generated by a small amount of growing ETEC is neutralized, so that the generation of the diarrhea is controlled from respective layers; the vaccine can not be limited by domains; the use range is relatively wide; and the protection rate is relatively high. After the vaccine is used, the protection rate is up to 95 percent; the immune efficiency of the vaccine is 5 times higher than that of the vaccine used in the market; and the recombined enterotoxin (namely, the fimbriae composite polyvalent vaccine) is safe and reliable.

Description

technical field [0001] The invention relates to the preparation and application of an anti-piglet diarrhea recombinant enterotoxin and pilus composite multivalent vaccine, which belongs to the technical field of animal husbandry and veterinary medicine. Background technique [0002] Diarrhea is caused by certain specific serotypes of enterotoxigenic E. coli (ETEC), which accounts for a large proportion of the deaths of newborn piglets and seriously endangers the pig industry. Healthy development, and caused huge economic losses to farmers every year. Diarrhea in piglets is mostly acute and highly contagious. The clinical symptoms are discharge of highly dilute yellow stool, which turns watery in severe cases, which in turn leads to dehydration and death of piglets. [0003] The pathogenic factors leading to pathogenic ETEC in piglet diarrhea are divided into two categories: adhesin and enterotoxin. Existing studies have shown that the types of ETEC pili that can cause diar...

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Application Information

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IPC IPC(8): A61K39/116A61P31/04A61K39/108
Inventor 徐永平张恒慧金礼吉尤建嵩李晓宇
Owner DALIAN UNIV OF TECH
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