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Porcine circovirus type 2 strain, inactivated vaccine prepared from porcine circovirus type 2 strain and application of porcine circovirus type 2 strain

A technology of porcine circovirus and virus liquid, which is applied in antiviral agents, virus/bacteriophage, virus antigen components, etc., can solve the problem of lack of PCV2 inactivated vaccine, etc., and achieve the effect of strong immune protection and good immunogenicity

Active Publication Date: 2015-11-25
JILIN ZHENGYE BIOLOGICAL PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there is still a lack of a safe, efficient and cheap PCV2 inactivated vaccine

Method used

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  • Porcine circovirus type 2 strain, inactivated vaccine prepared from porcine circovirus type 2 strain and application of porcine circovirus type 2 strain
  • Porcine circovirus type 2 strain, inactivated vaccine prepared from porcine circovirus type 2 strain and application of porcine circovirus type 2 strain
  • Porcine circovirus type 2 strain, inactivated vaccine prepared from porcine circovirus type 2 strain and application of porcine circovirus type 2 strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Isolation and Identification of Porcine Circovirus Type 2 Yh-1 Strain

[0035] 1. Experimental method

[0036] 1.1 Disease samples and cells for virus isolation

[0037]The lungs and lymph nodes of the dead piglets in a pig farm in Inner Mongolia were collected and stored in a -70°C ultra-low temperature freezer for later use. PK-15 cells are preserved by our research and development center, using GibcoMEM medium containing 10% calf serum and 5% CO at 37°C 2 Incubator cultivation.

[0038] 1.2 Treatment of disease materials

[0039] Cut a certain volume of disease material, add 5 times the volume of MEM with 5% serum, grind it with a glass grinder, centrifuge the homogenate at 12000rpm at 4°C for 10min, and filter the supernatant with a 0.22μm filter for sterilization before use.

[0040] 1.3 Primer design

[0041] PCV2-specific detection primers were designed according to the published PCV2 sequence, the virus gene sequence was amplified by PCR method, s...

Embodiment 2

[0060] The preparation of the inactivated vaccine of embodiment 2 porcine circovirus type 2 Yh-1 strains

[0061] 1. Experimental method

[0062] 1.1 Virus inactivation

[0063] Add 0.2% of the total amount of PCV-2Yh-1 strain F15 virus solution into formaldehyde solution, shake for 5 minutes, then inactivate at 37°C for 20 hours, shake 3 to 4 times, 3 minutes each time. Store the inactivation solution at 2-8°C.

[0064] 1.2 Inactivation test

[0065] Take the inactivated virus solution of PCV-2Yh-1 strain, inoculate pk-15 cells that have grown into a monolayer by 50 times dilution, inoculate 4 bottles of each sample, 1 mL per bottle, after adsorption for 1 hour, add maintenance solution to the original volume at 37°C Continue to culture for 4 days, there should be no cytopathic changes; continue to culture, and then blind passage for 2 generations, and then use indirect immunofluorescence and PCR to detect the virus. At the same time, the same batch of uninactivated virus...

experiment example 2

[0086] Experimental Example 2 Safety Test of Porcine Circovirus Type 2 Inactivated Vaccine

[0087] Fifteen PRRSV-negative piglets aged 4-5 weeks were randomly divided into 3 groups, 5 piglets / group, group I received a single dose of inactivated vaccine, group II received a double dose of inactivated vaccine, and group III was injected with emulsified MEM medium 2 mL was used as a control. Measure the body temperature before immunization, and measure the body temperature every day for 7 days after immunization; observe the feeding situation of the pigs for 14 days after immunization, touch the local area, and see if there is any lump. 28 days after immunization, 2 pigs in each group were dissected to observe the absorption of the vaccine.

[0088]The body temperature of pigs before and after immunization with single dose and double dose was measured. There was no phenomenon of elevated body temperature in pigs before and after immunization. The pigs are fed and mentally nor...

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Abstract

The invention discloses a porcine circovirus type 2 strain, an inactivated vaccine prepared from the porcine circovirus type 2 strain and application of the porcine circovirus type 2 strain, and belongs to the fields of isolation and application of the porcine circovirus type 2 strain. The invention firstly discloses a porcine circovirus type 2 Yh-1 strain, wherein the microorganism preservation number of the strain is CGMCC No. 10409. The invention also discloses a method for preparing a porcine circovirus preventing inactivated strain by virtue of the Yh-1 strain, wherein the method comprises the following steps: (1) amplifying the porcine circovirus type 2 Yh-1 strain and collecting a virus liquid; (2) adding an inactivating agent to inactivate the virus liquid and concentrating the virus liquid; (3) preparing an aqueous phase and an oil phase; and (4) mixing the aqueous phase and the oil phase, and emulsifying so as to obtain the Yh-1 strain. Results of animal experiments show that the inactivated vaccine prepared from the porcine circovirus type 2 Yh-1 disclosed by the invention is high in safety and is good in immunogenicity, and an immune protective rate reaches 100%; and the inactivated vaccine can offer complete protection over circovirus type 2 strong virulence attack and can be used for effectively preventing and controlling epidemic porcine circovirus.

Description

technical field [0001] The present invention relates to porcine circovirus strains, in particular to a porcine circovirus type 2 strain, and the invention also relates to an inactivated vaccine prepared from the porcine circovirus type 2 strain and its application, belonging to porcine circovirus Isolation and application fields of virus type 2 strains. Background technique [0002] Porcine circovirus type 2 (PCV2) infection is the main cause of weaned piglet multisystem wasting syndrome (PMWS), which is characterized by immunosuppression and multisystem failure in weaned piglets. The diseased pigs mainly show poor growth and development, anemia , dyspnea, diarrhea, weakness, slow weight gain, swollen lymph nodes and other clinical symptoms. At the same time, because it is an immunosuppressive disease, the immune defense function of the body is reduced, so that mixed infections often occur clinically, resulting in so-called various syndromes, such as : Porcine dermatitis ne...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/12A61P31/20
Inventor 何玉友冯建民袁立民颜彤王国辉王石刘斌杨键李长艳廉维杨泽彬冯会柳志光白杨吴艳丽田丽华于志云曲红李海
Owner JILIN ZHENGYE BIOLOGICAL PROD
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