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Bivalent inactivated vaccine for porcine parvovirus and porcine epizootic diarrhea

A dual inactivated vaccine and porcine epidemic diarrhea technology, which is applied in the direction of vaccines, viruses, antiviral agents, etc., can solve the problems of poor clinical application effect and no effective prevention of porcine parvovirus, etc., to achieve resistance to attack and maintain The effect of long time and high antibody titer

Active Publication Date: 2016-08-24
YEBIO BIOENG OF QINGDAO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, foreign countries mainly use attenuated vaccines from South Korea, and domestically mainly use porcine transmissible gastroenteritis and other vaccines developed by Harbin Animal Research Institute, but the clinical application effect is not good
At present, there is no effective dual inactivated vaccine against porcine parvovirus and porcine epidemic diarrhea in the market

Method used

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  • Bivalent inactivated vaccine for porcine parvovirus and porcine epizootic diarrhea
  • Bivalent inactivated vaccine for porcine parvovirus and porcine epizootic diarrhea
  • Bivalent inactivated vaccine for porcine parvovirus and porcine epizootic diarrhea

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Embodiment 1: Screening of porcine parvovirus (PPV) VP2 gene

[0016] 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.

[0017] 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.

[0018] According to the antigenic characteristics and amino acid sequence of porcine parvovirus VP2 protein, a pair of pr...

Embodiment 2

[0020] Example 2: Preparation of recombinant porcine parvovirus VP2 protein

[0021] 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 identified correctly by PCR using primers primer 1 and primer 2...

Embodiment 3

[0030] Embodiment 3: the preparation of subunit inactivated vaccine

[0031] 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).

[0032] (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.

[0033] (2) Preparation of aqueous phase The porcine parvovirus VP2 protein was diluted with physiological saline to 150 μg / 0.1 ml. 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.

[0034](3) Emulsification Take 2 parts of the oil phase and put it in a high-speed s...

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Abstract

The invention provides bivalent inactivated vaccine for porcine parvovirus and porcine epizootic diarrhea. The bivalent inactivated vaccine comprises antigen and a vaccine adjuvant, wherein the antigen is a porcine epidemic diarrhea virus with the preservation number being CGMCC No. 8503 and porcine parvovirus VP2 protein with the preservation number being CCTCC M 2016098, wherein the porcine parvovirus VP2 protein is expressed by an X33-VP2 strain; the strain was preserved in China Center for Type Culture Collection on 9, March, 2016. The bivalent inactivated vaccine for porcine parvovirus and porcine epizootic diarrhea is good in immunogenicity, an immunized antibody is generated quickly, the generated antibody is high in titer and long in holding time and retention period, the immunizing dose is small, the selected vaccine adjuvant is easy to inject, and the two types of diseases can be prevented and treated through single injection; if immune injection is conducted before mating, piglets born by a pregnant sow can obtain good passive immunity, the piglets can generate high immunity and can resist to attack of powerful viruses, and the survival rate of the piglets is raised.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a dual inactivated vaccine of porcine parvovirus and porcine epidemic diarrhea. 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 p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/23A61K39/215A61P31/14A61P31/20
CPCA61K39/12A61K2039/5252A61K2039/552A61K2039/55505A61K2039/70C12N2750/14034C12N2770/20034A61K2300/00
Inventor 刘新文邹敏王秀丽郭伟伟宫晓王龙邹桂荣
Owner YEBIO BIOENG OF QINGDAO
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