A triple vaccine against porcine parvovirus, porcine epidemic diarrhea and Escherichia coli

A technology of porcine epidemic diarrhea and triple vaccine, applied in the field of veterinary biological products, can solve the problem of ineffective prevention of porcine parvovirus, and achieve the effect of resisting attack, fast antibody production and good immunogenicity

Active Publication Date: 2021-02-05
YEBIO BIOENG OF QINGDAO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no effective triple vaccine against porcine parvovirus, porcine epidemic diarrhea, and Escherichia coli in the market at present.

Method used

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  • A triple vaccine against porcine parvovirus, porcine epidemic diarrhea and Escherichia coli
  • A triple vaccine against porcine parvovirus, porcine epidemic diarrhea and Escherichia coli
  • A triple vaccine against porcine parvovirus, porcine epidemic diarrhea and Escherichia coli

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Embodiment 1: Screening of porcine parvovirus (PPV) VP2 gene

[0017] In 2010, symptoms of porcine parvovirus disease appeared in many farms in Shandong Province, and the individual pigs with the disease had been injected with the existing parvovirus vaccine before, and it was speculated that the infected virus had mutated; Screening; the porcine parvovirus PPV1 was finally screened out.

[0018] In order to verify the antigenicity of the screened virus, five virus strains from different sources, including the screened PPV1 strain, were used as antigens to prepare vaccines. After immunizing SPF pigs, the screened virus liquid was used to challenge the virus. The results showed that Compared with other porcine parvovirus vaccines, the vaccine prepared by itself has a better immune effect (p<0.05), so it is confirmed that it has a genetic variation.

[0019] According to the antigenic characteristics and amino acid sequence of porcine parvovirus VP2 protein, a pair of pr...

Embodiment 2

[0021] Example 2: Preparation of recombinant porcine parvovirus VP2 protein

[0022] The method comprises the following steps: a. constructing an expression vector; b. constructing an expression strain; c. inducing, extracting and purifying the recombinant VP2 protein. The details are as follows: The positive cloned plasmid pMD18-T-VP2 and the expression vector pPICZα vector were digested with KpnI and NotI respectively, and after 1.2% agarose gel electrophoresis, they were recovered with a DNA gel recovery kit, and about 1.7 kb were obtained respectively. and 3.3kb fragments were oriented at 16°C to construct pPICZα-VP2 expression vector, after the correct identification by restriction enzyme digestion; after the plasmid was linearized, it was electrotransformed into Pichia pastoris competent cells to construct Pichia pastoris expression strain X33-VP2, The genome of the expressed strain X33-VP2 was extracted, and the PCR identification using primers primer 1 and primer 2 was...

Embodiment 3

[0031] Embodiment 3: the preparation of subunit inactivated vaccine

[0032] The semi-finished VP2 protein antigen after passing the inspection is used for vaccine preparation (the liquid components in the following preparations are calculated by volume ratio).

[0033] (1) Preparation of oil phase Take 95 parts of veterinary white oil and 1 part of aluminum stearate, put them in the oil phase preparation tank and heat to 80°C, then add 5 parts of Siben-80, until the temperature reaches 115°C, Keep it for 30 minutes, and set aside after cooling.

[0034] (2) Preparation of aqueous phase Porcine parvovirus VP2 protein was diluted to 150 μg / 0.1 ml with physiological saline. Take 5 parts of sterilized Tween-80 and add them to the liquid mixing tank, and at the same time add 95 parts of protein solution for seedling production, and stir for 20-30 minutes to completely dissolve the Tween-80.

[0035] (3) Emulsification Take 2 parts of the oil phase and put it in a high-speed sheari...

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Abstract

The invention provides a porcine parvovirus, porcine epidemic diarrhea and Escherichia coli triple vaccine, which contains an antigen and a vaccine adjuvant, wherein the antigen is porcine parvovirus VP2 expressed by the X33-VP2 strain with the preservation number CCTCC M 2016098 Protein; ciliary antigens of porcine epidemic diarrhea virus and Escherichia coli K88, K99, 987P with preservation number CGMCC No.8503. The porcine parvovirus, porcine epidemic diarrhea and Escherichia coli triple vaccine of the present invention has good immunogenicity, rapid antibody production after immunization, high antibody titer and long maintenance time, long storage period, and small immunization dose. The adjuvant is easy to inject, and can prevent and treat three diseases with one injection. Immunization injection before mating can make piglets born by pregnant sows get better passive immunity, so that piglets can develop strong immunity and can resist strong poisonous attacks. Improve the survival rate of piglets.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, in particular to a triple vaccine of porcine parvovirus, porcine epidemic diarrhea and Escherichia coli. Background technique [0002] Porcine parvovirus (PPV) is one of the important pathogens causing reproductive disorders in sows, which can lead to abortion, premature birth, stillbirth, mummified fetuses, weak piglets, infertility in sows and mass death of newborn piglets. Since the 1980s in my country, PPV has been isolated from various places, and the survey found that the serological positive rate of the disease in some pig herds can reach more than 85%. PPV often presents as recessive infection, especially the phenomenon of low-dose persistent infection often occurs. In addition to causing reproductive disorders in sows, the virus can also cooperate with other pathogens to cause a variety of diseases, causing great economic losses to the pig industry. Porcine parvo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/23A61K39/108A61K39/215A61P31/04A61P1/12
CPCA61K39/0258A61K39/12A61K2039/545A61K2039/552A61K2039/55505A61K2039/70C12N2750/14334C12N2770/20034A61K2300/00
Inventor 刘新文杜元钊王秀丽郭玉广陶晓珊刘蕾
Owner YEBIO BIOENG OF QINGDAO
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